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Abstract:

Methods and pharmaceutical compositions for treating viral infections, by
administering certain thienopyridine derivative compounds in
therapeutically effective amounts are disclosed. Methods of using the
compounds and pharmaceutical compositions thereof are also disclosed. In
particular, the treatment of viral infections such as caused by
flavivirus is disclosed, i.e., including but not limited to, Dengue
virus, West Nile virus, yellow fever virus, Japanese encephalitis virus,
and tick-borne encephalitis virus.

38. The method of claim 37, wherein said compound is
3-amino-N,6-bis(4-chlorophenyl)thieno[2,3-b]pyridine-2-carboxamide.

39. The method of claim 37, wherein said compound is
3-amino-6-[3-(difluoromethoxy)phenyl]-N-[4-(difluoromethoxy)phenyl]thieno-
[2,3-b]pyridine-2-carboxamide.

40. The method of claim 37, wherein the mammal is a human.

41. The method of claim 37, wherein said Dengue virus is selected from
the group consisting of DEN-1, DEN-2, DEN-3, and DEN-4.

42. The method of claim 37, wherein said viral infection is associated
with Dengue fever.

43. The method of claim 42, wherein said Dengue fever is selected from
the group consisting of classical dengue fever and dengue hemorrhagic
fever.

44. The method of claim 37, which further comprises co-administration of
at least one agent selected from the group consisting of antiviral agent,
vaccine, and interferon.

45. The method of claim 44, wherein said interferon is pegylated.

46. A pharmaceutical composition comprising a pharmaceutically acceptable
carrier and a compound or a pharmaceutically acceptable salt thereof,
wherein said compound is selected from the group consisting of:
3-amino-N-(4-bromophenyl)-6-(4-chlorophenyl)thieno[2,3-b]pyridine-2-carbo-
xamide; 3-amino-6-(3-methoxyphenyl)-N-[4-(trifluoromethoxy)phenyl]thieno[2-
,3-b]pyridine-2-carboxamide;
3-amino-N-(2,5-dichlorophenyl)-6-(2-thienyl)thieno[2,3-b]pyridine-2-carbo-
xamide; 3-amino-N-(2,3-dichlorophenyl)-6-(2-thienyl)thieno[2,3-b]pyridine--
2-carboxamide;
3-amino-N-(4-bromophenyl)-6-(3-methoxyphenyl)thieno[2,3-b]pyridine-2-carb-
oxamide; 3-amino-6-(1,3-benzodioxol-5-yl)-N-(2-bromo-4-methyl-phenyl)thien-
o[2,3-b]pyridine-2-carboxamide; and
3-amino-6-(3-methoxyphenyl)-N-(2-phenoxyphenyl)thieno[2,3-b]pyridine-2-ca-
rboxamide, wherein said composition is suitable for human or animal
administration.

47. The composition of claim 46, wherein said compound is
3-amino-N-(4-bromophenyl)-6-(4-chlorophenyl)thieno[2,3-b]pyridine-2-carbo-
xamide.

48. The composition of claim 46, wherein said compound is
3-amino-6-(3-methoxyphenyl)-N-[4-(trifluoromethoxy)phenyl]thieno[2,3-b]py-
ridine-2-carboxamide.

49. A method for the treatment of at least one type of a Dengue virus
infection or disease associated therewith, comprising administering in a
therapeutically effective amount to a mammal in need thereof, a compound
or a pharmaceutically acceptable salt thereof, wherein said compound is
selected from the group consisting of:
3-amino-N-(4-bromophenyl)-6-(4-chlorophenyl)thieno[2,3-b]pyridine-2-carbo-
xamide; 3-amino-6-(3-methoxyphenyl)-N-[4-(trifluoromethoxy)phenyl]thieno[2-
,3-b]pyridine-2-carboxamide;
3-amino-N-(2,5-dichlorophenyl)-6-(2-thienyl)thieno[2,3-b]pyridine-2-carbo-
xamide; 3-amino-N-(2,3-dichlorophenyl)-6-(2-thienyl)thieno[2,3-b]pyridine--
2-carboxamide;
3-amino-N-(4-bromophenyl)-6-(3-methoxyphenyl)thieno[2,3-b]pyridine-2-carb-
oxamide; 3-amino-6-(1,3-benzodioxol-5-yl)-N-(2-bromo-4-methyl-phenyl)thien-
o[2,3-b]pyridine-2-carboxamide; and
3-amino-6-(3-methoxyphenyl)-N-(2-phenoxyphenyl)thieno[2,3-b]pyridine-2-ca-
rboxamide.

50. The method of claim 49, wherein said compound is
3-amino-N-(4-bromophenyl)-6-(4-chlorophenyl)thieno[2,3-b]pyridine-2-carbo-
xamide.

51. The method of claim 49, wherein said compound is
3-amino-6-(3-methoxyphenyl)-N-[4-(trifluoromethoxy)phenyl]thieno[2,3-b]py-
ridine-2-carboxamide.

52. The method of claim 49, wherein the mammal is a human.

53. The method of claim 49, wherein said Dengue virus is selected from
the group consisting of DEN-1, DEN-2, DEN-3, and DEN-4.

54. The method of claim 53, wherein said viral infection is associated
with Dengue fever.

55. The method of claim 54, wherein said Dengue fever is selected from
the group consisting of classical dengue fever and dengue hemorrhagic
fever.

56. The method of claim 49, which further comprises co-administration of
at least one agent selected from the group consisting of antiviral agent,
vaccine, and interferon.

57. The method of claim 56, wherein said interferon is pegylated.

Description:

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation-in-part and claims priority to
U.S. patent application Ser. No. 13/203,351, filed Oct. 13, 2011, which
is a national stage entry under U.S.C. 371(c), and claims priority to
International Patent Application Number PCT/US10/25183, filed Feb. 24,
2010, which in turn claims priority to and benefit of U.S. Provisional
Application No. 61/156,132, filed Feb. 27, 2009. All the applications are
incorporated herein by reference in the entirety and for all purposes.

FIELD OF THE INVENTION

[0003] This invention relates to the use of thienopyridine derivatives and
analogs, as well as compositions containing the same, for the treatment
of viral diseases associated with the flavivirus family such as Dengue
fever, Yellow fever, West Nile, St. Louis encephalitis, Hepatitis C,
Murray Valley encephalitis, and Japanese encephalitis.

BACKGROUND OF THE INVENTION

[0004] Dengue fever (DF) is an acute febrile disease caused by one of four
closely related virus serotypes (DEN-1, DEN-2, DEN-3, and DEN-4). Dengue
fever is classified based on its clinical characteristics into classical
dengue fever, or the more severe forms, dengue hemorrhagic fever syndrome
(DHF), and dengue shock syndrome (DSS). Recovery from infection from one
serotype produces life-long immunity to that particular serotype, but
provides only short-lived and limited protection against any of the other
serotypes (32). Dengue is a member of the Flaviviridae family which are
enveloped, positive-sense RNA viruses whose human pathogens also include
West Nile virus (WNV), yellow fever virus (YFV), Japanese encephalitis
virus (JEV), and tick-borne encephalitis virus (TBEV) among others.
Dengue transmission is via the bite of an infected Aedes aegypti mosquito
which is found in tropical and sub-tropical regions around the world.

[0005] Each year regional epidemics of dengue cause significant morbidity
and mortality, social disruption and substantial economic burden on the
societies affected both in terms of hospitalization and mosquito control.
Dengue is considered by the World Health Organization (WHO) to be the
most important arthropod-borne viral disease with an estimated 50 million
cases of dengue infection, including 500,000 DHF cases and 24,000 deaths
worldwide each year (32, 33). WHO estimates that forty percent of the
world's population (2.5 billion people) are at risk for DF, DHF, and DSS
(32). Dengue is also a NIAID Category A pathogen and in terms of
bio-defense, represents a significant threat to United States troops
overseas. Dengue is an emerging threat to North America with a dramatic
increase in severe disease in the past 25 years including major epidemics
in Cuba and Venezuela, and outbreaks in Texas and Hawaii (4). Failure to
control the mosquito vector and increases in long-distance travel have
contributed to the increase and spread of dengue disease. The
characteristics of dengue as a viral hemorrhagic fever virus
(arthropod-borne, widely spread, and capable of inducing a great amount
of cellular damage and eliciting an immune response that can result in
severe hemorrhage, shock, and death) makes this virus a unique threat to
deployed military personnel around the world as well as to travelers to
tropical regions. Preparedness for both biodefense and for the public
health challenges posed by dengue will require the development of new
vaccines and antiviral therapeutics.

[0006] Dengue causes several illnesses with increasing severity being
determined in part by prior infection with a different serotype of the
virus. Classic dengue fever (DF) begins 3-8 days after the bite of an
infected mosquito and is characterized by sudden onset of fever,
headache, back pain, joint pain, a measles-like rash, and nausea and
vomiting (20). DF is frequently referred to as "breakbone" fever due to
these symptoms. The disease usually resolves after two weeks but a
prolonged recovery with weakness and depression is common. The more
severe form of the disease, dengue hemorrhagic fever (DHF) has a similar
onset and early phase of illness as dengue fever. However, shortly after
onset the disease is characterized by high fever, enlargement of the
liver, and hemorrhagic phenomena such as bleeding from the nose, mouth,
and internal organs due to vascular permeability (33). In dengue shock
syndrome (DSS) circulatory failure and hypovolaemic shock resulting from
plasma leakage occur and can lead to death in 12-24 hours without plasma
replacement (33). The case fatality rate of DHF/DSS can be as high as 20%
without treatment. DHF has become a leading cause of hospitalization and
death among children in many countries with an estimated 500,000 cases
requiring hospitalization each year and a case fatality rate of about 5%
(32).

[0007] The pathogenesis of DHF/DSS is still being studied but is thought
to be due in part to an enhancement of virus replication in macrophages
by heterotypic antibodies, termed antibody-dependent enhancement (ADE)
(8). During a secondary infection, with a different serotype of dengue
virus, cross-reactive antibodies that are not neutralizing form
virus-antibody complexes that are taken into monocytes and Langerhans
cells (dendritic cells) and increase the number of infected cells (7).
This leads to the activation of cytotoxic lymphocytes which can result in
plasma leakage and the hemorrhagic features characteristic of DHF and DSS
(20). This antibody-dependent enhancement of infection is one reason why
the development of a successful vaccine has proven to be so difficult.
Although less frequent, DHF/DSS can occur after primary infection (29),
so virus virulence (15) and immune activation are also believed to
contribute to the pathogenesis of the disease (25).

[0008] Dengue is endemic in more than 100 countries in Africa, the
Americas, the Eastern Mediterranean, South-east Asia and the Western
Pacific. During epidemics, attack rates can be as high as 80-90% of the
susceptible population. All four serotypes of the virus are emerging
worldwide, increasing the number of cases of the disease as well as the
number of explosive outbreaks. In 2002 for example, there were 1,015,420
reported cases of dengue in the Americas alone with 14,374 cases of DHF,
which is more than three times the number of dengue cases reported in the
Americas in 1995 (23).

[0009] The dengue genome, approximately 11 kb in length, consists of a
linear, single stranded, infectious, positive sense RNA that is
translated as a single long polyprotein (reviewed in (27)). The genome is
composed of seven nonstructural (NS) protein genes and three structural
protein genes which encode the nucleocapsid protein (C), a
membrane-associated protein (M), and an envelope protein (E). The
nonstructural proteins are involved in viral RNA replication (31), viral
assembly, and the inflammatory components of the disease (18). The
structural proteins are involved mainly in viral particle formation (21).
The precursor polyprotein is cleaved by cellular proteinases to separate
the structural proteins (17), while a virus-encoded proteinase cleaves
the nonstructural region of the polyprotein (6). The genome is capped and
does not have a poly(A) tail at the 3' end but instead has a stable
stem-loop structure necessary for stability and replication of the
genomic RNA (3). The virus binds to cellular receptors via the E protein
and undergoes receptor-mediated endocytosis followed by low-pH fusion in
lysosomes (19). The viral genome is then uncoated and translated into the
viral precursor polyprotein. Co- and posttranslational proteolytic
processing separates the structural and nonstructural proteins. The
RNA-dependent RNA polymerase along with cofactors synthesizes the
minus-strand RNA which serves as a template for the synthesis of the
progeny plus-strand RNA (24). Viral replication is membrane associated
(1, 30). Following replication, the genome is encapsidated, and the
immature virus, surrounded by a lipid envelope buds into the lumen (9).
The envelope proteins become glycosylated and mature viruses are released
outside the cell. Essential stages or process during the virus life cycle
would be possible targets for inhibition from an antiviral drug and
include binding of the virus to the cell through the E protein, uptake of
the virus into the cell, the capping mechanism, the viral proteinase, the
viral RNA-dependent RNA polymerase, and the viral helicase.

[0010] Current management of dengue virus-related disease relies solely on
vector control. There are no approved antivirals or vaccines for the
treatment or prevention of dengue. Ribavirin, a guanosine analogue, has
been shown to be effective against a range of RNA virus infections and
works against dengue in tissue culture by inhibiting the dengue
2'-O-methyltransferase NS5 domain (2, 10). However, ribavirin did not
show protection against dengue in a mouse model (14) or a rhesus monkey
model (16), instead it induced anemia and thrombocytosis. While there are
no currently available approved vaccines, multivalent dengue vaccines
have shown some limited potential in humans (5, 11, 12, 26). However,
vaccine development is difficult due to the presence of four distinct
serotypes of the virus which each cause disease. Vaccine development also
faces the challenge of ADE where unequal protection against the different
strains of the virus could actually increase the risk of more serious
disease. Therefore there is a need for antiviral drugs that target all of
the serotypes of dengue. An antiviral drug administered early during
dengue infection that inhibits viral replication would prevent the high
viral load associated with DHF and be an attractive strategy in the
treatment and prevention of disease. An antiviral drug that inhibits
viral replication could be administered prior to travel to a dengue
endemic region to prevent acquisition of disease, or for those that have
previously been exposed to dengue, could prevent infection by another
serotype of virus and decrease the chance of life-threatening DHF and
DSS. Having an antiviral drug would also aid vaccine development by
having a tool at hand to treat complications that may arise due to
unequal immune protection against the different serotypes. Although a
successful vaccine could be a critical component of an effective
biodefense, the typical delay to onset of immunity, potential
side-effects, cost, and logistics associated with large-scale civilian
vaccinations against a low-threat risk agent suggest that a comprehensive
biodefense include a separate rapid-response element. Thus, there remains
an urgent need to develop a safe and effective product to protect against
flavivirus infection.

SUMMARY OF THE INVENTION

[0011] The present invention provides a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound having
the following general Formula I or a pharmaceutically acceptable salt
thereof:

[0013] R is selected from the group consisting of halogen, cyano,
isocyano, nitro, amino, alkylamino, dialkylamino, cycloalkylamino,
heterocycloalkylamino, arylamino, heteroarylamino, acylamino,
arylacylamino, heteroarylacylamino, alkylsulfonylamino,
arylsulfonylamino, hydroxysulfonyl, aminosulfonyl, substituted
aminosulfonyl, acyl, arylacyl, heteroarylacyl, carboxy, alkoxycarbonyl,
cycloalkyloxycarbonyl, aryloxycarbonyl, aminocarbonyl, and substituted
aminocarbonyl, or R and R1 together with the carbons they are
attached to may form a substituted or unsubstituted ring; and

[0014] A, B, D, and E are independently N or C--R1, C--R2,
C--R3 and C--R4, respectively, wherein R1, R2,
R3 and R4 are independently selected from the group consisting
of hydrogen, substituted or unsubstituted alkyl, alkenyl, alkynyl,
cycloalkyl, heterocycloalkyl, arylalkyl, aryl, heteroaryl, hydroxy,
alkyloxy, aryloxy, heteroaryloxy, acyloxy, arylacyloxy,
heteroarylacyloxy, alkylsulfonyloxy, arylsulfonyloxy, thio, alkylthio,
arylthio, amino, alkylamino, dialkylamino, cycloalkylamino,
heterocycloalkylamino, arylamino, heteroarylamino, acylamino,
arylacylamino, heteroarylacylamino, alkylsulfonylamino,
arylsulfonylamino, acyl, arylacyl, heteroarylacyl, alkylsulfinyl,
arylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, substituted
aminosulfonyl, carboxy, alkoxycarbonyl, cycloalkyloxycarbonyl,
aryloxycarbonyl, carbamoyl, substituted carbamoyl, halogen, cyano,
isocyano and nitro; or R1 and R together with the carbons they are
attached to may form a substituted or unsubstituted ring, or R2 and
R3 or R3 and R4 together with the carbons they are
attached to may form a substituted or unsubstituted ring, which may be
aromatic or non-aromatic and may include one or more heteroatoms in the
ring and may be fused with an aromatic or aliphatic ring. The
pharmaceutical composition must be suitable for human or animal
administration.

[0015] The present invention also provides a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound having
the following general Formula II or a pharmaceutically acceptable salt
thereof:

[0018] G is selected from the group consisting of --C(═O)--,
--C(═S)--, --S(═O)2--, and --C(═NR5)--, wherein
R5 is selected from the groups consisting of hydrogen, alkyl,
alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, arylalkyl, aryl,
heteroaryl, acyl, arylacyl, heteroarylacyl, sulfonyl, aminosulfonyl,
substituted aminosulfonyl, alkoxycarbonyl, cycloalkyloxycarbonyl,
aryloxycarbonyl, carbamoyl and substituted carbamoyl; or R5 and
R6 or R7, together with the nitrogen atoms they are attached
to, along with the carbon of G, or R5 and R8 or R9,
together with the nitrogen atoms they are attached to, along with the
carbon of G and two carbons of the X-containing 5-membered ring, may form
a substituted or unsubstituted ring, which may be fused with an aromatic
or aliphatic ring;

[0019] R6, R7, R8, and R9 are independently selected
from the groups consisting of hydrogen, alkyl, alkenyl, alkynyl,
cycloalkyl, heterocycloalkyl, arylalkyl, aryl, heteroaryl, acyl,
arylacyl, heteroarylacyl, sulfonyl, aminosulfonyl, substituted
aminosulfonyl, alkoxycarbonyl, cycloalkyloxycarbonyl, aryloxycarbonyl,
carbamoyl and substituted carbamoyl; or R6 or R7 and R5,
together with the nitrogen atoms they are attached to, along with the
carbon of G, or R8 or R9 and R5, together with the
nitrogen atoms they are attached to, along with the carbon of G and two
carbons of the X-containing 5-membered ring, or R6 or R7 and
R8 or R9, together with the nitrogen atoms they are attached
to, along with the carbon or sulfur of G and two carbons of the
X-containing 5-membered ring, or R6 and R7, together with the
nitrogen atom they are attached to, or R8 and R9, together with
the nitrogen atom they are attached to, may form a substituted or
unsubstituted ring, which may be fused with an aromatic or aliphatic
ring; and

##STR00003##

is a 7 or 8-membered ring which contains one or more heteroatoms selected
from N, O and S, or a 4-membered ring which may optionally contain one or
more heteroatoms selected from N, O and S. The ring may be substituted or
unsubstituted, or fused with another ring, which may be aromatic or
non-aromatic and may include one or more heteroatoms in the ring and may
be fused with an aromatic or aliphatic ring. The pharmaceutical
composition must be suitable for human or animal administration.

[0020] The present invention further provides a pharmaceutical composition
comprising a pharmaceutically acceptable carrier and a compound having
the following general Formula III or a pharmaceutically acceptable salt
thereof:

[0023] B, D, and E are independently N or C--R2, C--R3 and
C--R4, respectively, wherein R2, R3 and R4 are
independently selected from the group consisting of hydrogen, substituted
or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,
arylalkyl, aryl, heteroaryl, hydroxy, alkyloxy, aryloxy, heteroaryloxy,
acyloxy, arylacyloxy, heteroarylacyloxy, alkylsulfonyloxy,
arylsulfonyloxy, thio, alkylthio, arylthio, amino, alkylamino,
dialkylamino, cycloalkylamino, heterocycloalkylamino, arylamino,
heteroarylamino, acylamino, arylacylamino, heteroarylacylamino,
alkylsulfonylamino, arylsulfonylamino, acyl, arylacyl, heteroarylacyl,
alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,
substituted aminosulfonyl, carboxy, alkoxycarbonyl,
cycloalkyloxycarbonyl, aryloxycarbonyl, carbamoyl, substituted carbamoyl,
halogen, cyano, isocyano and nitro; or R2 and R3 or R3 and
R4 together with the carbons they are attached to may form a
substituted or unsubstituted ring, which may be aromatic or non-aromatic
and may include one or more heteroatoms in the ring and may be fused with
an aromatic or aliphatic ring; and

[0031] G is selected from the group consisting of --C(═O)--,
--C(═S)--, --S(═O)2--, and --C(═NR5)--, wherein
R5 is selected from the groups consisting of hydrogen, alkyl,
alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, arylalkyl, aryl,
heteroaryl, acyl, arylacyl, heteroarylacyl, sulfonyl, aminosulfonyl,
substituted aminosulfonyl, alkoxycarbonyl, cycloalkyloxycarbonyl,
aryloxycarbonyl, carbamoyl and substituted carbamoyl; or R5 and
R6 or R7, together with the nitrogen atoms they are attached
to, along with the carbon of G, or R5 and R8 or R9,
together with the nitrogen atoms they are attached to, along with the
carbon of G and two carbons of the X-containing 5-membered ring, may form
a substituted or unsubstituted ring, which may be fused with an aromatic
or aliphatic ring;

[0032] R6, R7, R8, and R9 are independently selected
from the groups consisting of hydrogen, alkyl, alkenyl, alkynyl,
cycloalkyl, heterocycloalkyl, arylalkyl, aryl, heteroaryl, acyl,
arylacyl, heteroarylacyl, sulfonyl, aminosulfonyl, substituted
aminosulfonyl, alkoxycarbonyl, cycloalkyloxycarbonyl, aryloxycarbonyl,
carbamoyl and substituted carbamoyl; or R6 or R7 and R5,
together with the nitrogen atoms they are attached to, along with the
carbon of G, or R8 or R9 and R5, together with the
nitrogen atoms they are attached to, along with the carbon of G and two
carbons of the X-containing 5-membered ring, or R6 or R7 and
R8 or R9, together with the nitrogen atoms they are attached
to, along with the carbon or sulfur of G and two carbons of the
X-containing 5-membered ring, or R6 and R7, together with the
nitrogen atom they are attached to, or R8 and R9, together with
the nitrogen atom they are attached to, may form a substituted or
unsubstituted ring, which may be fused with an aromatic or aliphatic
ring; and

##STR00006##

is a 7 or 8-membered ring which contains one or more heteroatoms selected
from N, O and S, or a 4-membered ring which may optionally contain one or
more heteroatoms selected from N, O and S. The ring may be substituted or
unsubstituted, or fused with another ring, which may be aromatic or
non-aromatic and may include one or more heteroatoms in the ring and may
be fused with an aromatic or aliphatic ring.

[0033] The present invention also provides a compound having the following
general Formula III or a pharmaceutically acceptable salt thereof:

[0036] B, D, and E are independently N or C--R2, C--R3 and
C--R4, respectively, wherein R2, R3 and R4 are
independently selected from the group consisting of hydrogen, substituted
or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,
arylalkyl, aryl, heteroaryl, hydroxy, alkyloxy, aryloxy, heteroaryloxy,
acyloxy, arylacyloxy, heteroarylacyloxy, alkylsulfonyloxy,
arylsulfonyloxy, thio, alkylthio, arylthio, amino, alkylamino,
dialkylamino, cycloalkylamino, heterocycloalkylamino, arylamino,
heteroarylamino, acylamino, arylacylamino, heteroarylacylamino,
alkylsulfonylamino, arylsulfonylamino, acyl, arylacyl, heteroarylacyl,
alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,
substituted aminosulfonyl, carboxy, alkoxycarbonyl,
cycloalkyloxycarbonyl, aryloxycarbonyl, carbamoyl, substituted carbamoyl,
halogen, cyano, isocyano and nitro; or R2 and R3 or R3 and
R4 together with the carbons they are attached to may form a
substituted or unsubstituted ring, which may be aromatic or non-aromatic
and may include one or more heteroatoms in the ring and may be fused with
an aromatic or aliphatic ring; and

[0039] The present invention further provides a method for the treatment
of at least one type of a Dengue virus infection or disease associated
therewith, comprising administering in a therapeutically effective amount
to a mammal in need thereof, a compound of Formula I, II or III as
indicated above or a pharmaceutically acceptable salt thereof.

[0040] The present invention also provides a method for the treatment of
at least one type of a Dengue infection or disease associated therewith,
comprising administering in a therapeutically effective amount to a
mammal in need thereof, a compound or a pharmaceutically acceptable salt
thereof, wherein said compound is selected from the group consisting of:
3-amino-N-(4-bromophenyl)-6-(4-chlorophenyl)thieno[2,3-b]pyridine-2-carbo-
xamide; 3-amino-6-(3-methoxyphenyl)-N-[4-(trifluoromethoxy)phenyl]thieno[2-
,3-b]pyridine-2-carboxamide;
3-amino-N-(2,5-dichlorophenyl)-6-(2-thienyl)thieno[2,3-b]pyridine-2-carbo-
xamide; 3-amino-N-(2,3-dichlorophenyl)-6-(2-thienyl)thieno[2,3-b]pyridine--
2-carboxamide;
3-amino-N-(4-bromophenyl)-6-(3-methoxyphenyl)thieno[2,3-b]pyridine-2-carb-
oxamide; 3-amino-6-(1,3-benzodioxol-5-yl)-N-(2-bromo-4-methyl-phenyl)thien-
o[2,3-b]pyridine-2-carboxamide; and
3-amino-6-(3-methoxyphenyl)-N-(2-phenoxyphenyl)thieno[2,3-b]pyridine-2-ca-
rboxamide.

[0041] The present invention further provides novel intermediate compounds
used in the synthesis of the compounds of the present invention. These
intermediate compounds are selected from the group consisting of:
tert-butyl (4E)-4-(hydroxymethylene)-5-oxoazepane-1-carboxylate;
tert-butyl (3E)-3-(hydroxymethylene)-4-oxoazepane-1-carboxylate;
tert-butyl
3-cyano-2-thioxo-1,2,5,6,8,9-hexahydro-7H-pyrido[2,3-d]azepine-7-carboxyl-
ate; tert-butyl
3-cyano-2-thioxo-1,2,5,7,8,9-hexahydro-6H-pyrido[3,2-c]azepine-6-carboxyl-
ate; and 3-amino-7-tert-butyloxycarbonyl-6,7,8,9-tetrahydro-5H-1-thia-7,10-
-diaza-cyclohepta[f]indene-2-carboxylic acid
(5-phenyl-[1,3,4]thiadiazol-2-yl)-amide; and
3-amino-6-tert-butyloxycarbonyl-6,7,8,9-tetrahydro-5H-1-thia-6,10-diaza-c-
yclohepta[f]indene-2-carboxylic acid
(5-phenyl-[1,3,4]thiadiazol-2-yl)-amide.

[0042] The present invention further provides a method for the preparation
of a mixture of tert-butyl
(4E)-4-(hydroxymethylene)-5-oxoazepane-1-carboxylate and tert-butyl
(3E)-3-(hydroxymethylene)-4-oxoazepane-1-carboxylate, said method
comprising reacting tert-butyl 4-oxoazepane-1-carboxylate with
N-[tert-butoxy(dimethylamino)methyl]-N,N-dimethylamine.

[0043] The present invention also provides a method for the preparation of
a mixture of tert-butyl
3-cyano-2-thioxo-1,2,5,6,8,9-hexahydro-7H-pyrido[2,3-d]azepine-7-carboxyl-
ate and tert-butyl
3-cyano-2-thioxo-1,2,5,7,8,9-hexahydro-6H-pyrido[3,2-c]azepine-6-carboxyl-
ate said method comprising reacting a mixture of tert-butyl
(4E)-4-(hydroxymethylene)-5-oxoazepane-1-carboxylate and tert-butyl
(3E)-3-(hydroxymethylene)-4-oxoazepane-1-carboxylate in the presence of
2-cyanoethanethioamide and piperidine acetate.

[0044] The present invention further provides a method for the preparation
of 3-amino-7-tert-butyloxycarbonyl-6,7,8,9-tetrahydro-5H-1-thia-7,10-diaz-
a-cyclohepta[f]indene-2-carboxylic acid
(5-phenyl-[1,3,4]thiadiazol-2-yl)-amide comprising reacting tert-butyl
3-cyano-2-thioxo-1,2,5,6,8,9-hexahydro-7H-pyrido[2,3-d]azepine-7-carboxyl-
ate with 2-chloro-N-(5-phenyl-1,3,4-thiadiazol-2-yl)acetamide.

[0051] R is selected from the group consisting of halogen, cyano,
isocyano, nitro, amino, alkylamino, dialkylamino, cycloalkylamino,
heterocycloalkylamino, arylamino, heteroarylamino, acylamino,
arylacylamino, heteroarylacylamino, alkylsulfonylamino,
arylsulfonylamino, hydroxysulfonyl, aminosulfonyl, substituted
aminosulfonyl, acyl, arylacyl, heteroarylacyl, carboxy, alkoxycarbonyl,
cycloalkyloxycarbonyl, aryloxycarbonyl, aminocarbonyl, and substituted
aminocarbonyl, or R and R1 together with the carbons they are
attached to may form a substituted or unsubstituted ring; and

[0052] A, B, D, and E are independently N or C--R1, C--R2,
C--R3 and C--R4, respectively, wherein R1, R2,
R3 and R4 are independently selected from the group consisting
of hydrogen, substituted or unsubstituted alkyl, alkenyl, alkynyl,
cycloalkyl, heterocycloalkyl, arylalkyl, aryl, heteroaryl, hydroxy,
alkyloxy, aryloxy, heteroaryloxy, acyloxy, arylacyloxy,
heteroarylacyloxy, alkylsulfonyloxy, arylsulfonyloxy, thio, alkylthio,
arylthio, amino, alkylamino, dialkylamino, cycloalkylamino,
heterocycloalkylamino, arylamino, heteroarylamino, acylamino,
arylacylamino, heteroarylacylamino, alkylsulfonylamino,
arylsulfonylamino, acyl, arylacyl, heteroarylacyl, alkylsulfinyl,
arylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, substituted
aminosulfonyl, carboxy, alkoxycarbonyl, cycloalkyloxycarbonyl,
aryloxycarbonyl, carbamoyl, substituted carbamoyl, halogen, cyano,
isocyano and nitro; or R1 and R together with the carbons they are
attached to may form a substituted or unsubstituted ring, or R2 and
R3 or R3 and R4 together with the carbons they are
attached to may form a substituted or unsubstituted ring, which may be
aromatic or non-aromatic and may include one or more heteroatoms in the
ring and may be fused with an aromatic or aliphatic ring.

[0053] Preferably, for the compound of Formula I, X is S; A is
C--NH2, B is C--R2 and R2 is fluoro substituted phenyl or
B is C--H; D is a C--H; E is C--R4 and R4 is a thienyl or D is
C--R3 and E is C--R4, and R3 and R4 form a ring;
and/or R is a substituted aminocarbonyl.

[0055] More preferably, the compound of Formula I of the present invention
is 3-amino-6,7,8,9-tetrahydro-5H-1-thia-10-aza-cyclohepta[f]indene-2-carb-
oxylic acid (5-phenyl-[1,3,4]thiadiazol-2-yl)-amide.

[0056] The compounds of the invention are also of the following general
Formula II:

[0059] G is selected from the group consisting of --C(═O)--,
--C(═S)--, --S(═O)2--, and --C(═NR5)--, wherein
R5 is selected from the groups consisting of hydrogen, alkyl,
alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, arylalkyl, aryl,
heteroaryl, acyl, arylacyl, heteroarylacyl, sulfonyl, aminosulfonyl,
substituted aminosulfonyl, alkoxycarbonyl, cycloalkyloxycarbonyl,
aryloxycarbonyl, carbamoyl and substituted carbamoyl; or R5 and
R6 or R7, together with the nitrogen atoms they are attached
to, along with the carbon of G, or R5 and R8 or R9,
together with the nitrogen atoms they are attached to, along with the
carbon of G and two carbons of the X-containing 5-membered ring, may form
a substituted or unsubstituted ring, which may be fused with an aromatic
or aliphatic ring;

[0060] R6, R7, R8, and R9 are independently selected
from the groups consisting of hydrogen, alkyl, alkenyl, alkynyl,
cycloalkyl, heterocycloalkyl, arylalkyl, aryl, heteroaryl, acyl,
arylacyl, heteroarylacyl, sulfonyl, aminosulfonyl, substituted
aminosulfonyl, alkoxycarbonyl, cycloalkyloxycarbonyl, aryloxycarbonyl,
carbamoyl and substituted carbamoyl; or R6 or R7 and R5,
together with the nitrogen atoms they are attached to, along with the
carbon of G, or R8 or R9 and R5, together with the
nitrogen atoms they are attached to, along with the carbon of G and two
carbons of the X-containing 5-membered ring, or R6 or R7 and
R8 or R9, together with the nitrogen atoms they are attached
to, along with the carbon or sulfur of G and two carbons of the
X-containing 5-membered ring, or R6 and R7, together with the
nitrogen atom they are attached to, or R8 and R9, together with
the nitrogen atom they are attached to, may form a substituted or
unsubstituted ring, which may be fused with an aromatic or aliphatic
ring; and

##STR00010##

is a 7 or 8-membered ring which contains one or more heteroatoms selected
from N, O and S, or a 4-membered ring which may optionally contain one or
more heteroatoms selected from N, O and S. The ring may be substituted or
unsubstituted, or fused with another ring, which may be aromatic or
non-aromatic and may include one or more heteroatoms in the ring and may
be fused with an aromatic or aliphatic ring.

[0061] Preferably, for the compound of Formula II, X is S; B is CH; each
of R8 and R9 is H; G is --C(═O)--; R6 is a hydrogen;
R7 is a heteroaryl; and

##STR00011##

is a 7-membered ring which contains N as a heteroatom.

[0062] Preferably, the compound of Formula II of the present invention is
3-amino-6,7,8,9-tetrahydro-5H-1-thia-6,10-diaza-cyclohepta[f]indene-2-car-
boxylic acid (5-phenyl-[1,3,4]thiadiazol-2-yl)-amide.

[0063] Also preferably, the compound of Formula II of the present
invention is
3-amino-6,7,8,9-tetrahydro-5H-1-thia-7,10-diaza-cyclohepta[f]indene-2-car-
boxylic acid (5-phenyl-[1,3,4]thiadiazol-2-yl)-amide.

[0064] The compounds of the present invention are also of the following
general Formula III:

[0067] B, D, and E are independently N or C--R2, C--R3 and
C--R4, respectively, wherein R2, R3 and R4 are
independently selected from the group consisting of hydrogen, substituted
or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,
arylalkyl, aryl, heteroaryl, hydroxy, alkyloxy, aryloxy, heteroaryloxy,
acyloxy, arylacyloxy, heteroarylacyloxy, alkylsulfonyloxy,
arylsulfonyloxy, thio, alkylthio, arylthio, amino, alkylamino,
dialkylamino, cycloalkylamino, heterocycloalkylamino, arylamino,
heteroarylamino, acylamino, arylacylamino, heteroarylacylamino,
alkylsulfonylamino, arylsulfonylamino, acyl, arylacyl, heteroarylacyl,
alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl,
substituted aminosulfonyl, carboxy, alkoxycarbonyl,
cycloalkyloxycarbonyl, aryloxycarbonyl, carbamoyl, substituted carbamoyl,
halogen, cyano, isocyano and nitro; or R2 and R3 or R3 and
R4 together with the carbons they are attached to may form a
substituted or unsubstituted ring, which may be aromatic or non-aromatic
and may include one or more heteroatoms in the ring and may be fused with
an aromatic or aliphatic ring; and

[0069] Preferably, for the compound of Formula III, X is S; B is C--H; D
is C--H; and E is C--R4 and R4 is a heteroaryl. Also
preferably, D is C--R3 and E is C--R4, and R3 and R4
form a ring. Again preferably, R is a substituted aminocarbonyl.

[0070] Preferably, the compound of Formula III of the present invention is
3-[N-[3-amino-6-(2-thienyl)thieno[2,3-b]pyridine-2-carbonyl]-3-(trifluoro-
methyl)anilino]propanoic acid.

[0072] The compounds of the present invention also include a compound or a
pharmaceutically acceptable salt thereof, wherein said compound is
selected from the group consisting of:
3-amino-N-(4-bromophenyl)-6-(4-chlorophenyl)thieno[2,3-b]pyridine-2-carbo-
xamide; 3-amino-6-(3-methoxyphenyl)-N-[4-(trifluoromethoxy)phenyl]thieno[2-
,3-b]pyridine-2-carboxamide;
3-amino-N-(2,5-dichlorophenyl)-6-(2-thienyl)thieno[2,3-b]pyridine-2-carbo-
xamide; 3-amino-N-(2,3-dichlorophenyl)-6-(2-thienyl)thieno[2,3-b]pyridine--
2-carboxamide;
3-amino-N-(4-bromophenyl)-6-(3-methoxyphenyl)thieno[2,3-b]pyridine-2-carb-
oxamide; 3-amino-6-(1,3-benzodioxol-5-yl)-N-(2-bromo-4-methyl-phenyl)thien-
o[2,3-b]pyridine-2-carboxamide; and
3-amino-6-(3-methoxyphenyl)-N-(2-phenoxyphenyl)thieno[2,3-b]pyridine-2-ca-
rboxamide. Preferably said compound is
3-amino-N-(4-bromophenyl)-6-(4-chlorophenyl)thieno[2,3-b]pyridine-2-carbo-
xamide or 3-amino-6-(3-methoxyphenyl)-N-[4-(trifluoromethoxy)phenyl]thieno-
[2,3-b]pyridine-2-carboxamide.

[0073] The method of the present invention is for the treatment of at
least one type of a Dengue virus infection or disease associated
therewith (each type of Dengue virus infection being caused by a Dengue
virus serotype), comprising administering in a therapeutically effective
amount to a mammal in need thereof, a compound of Formula I, Formula II,
Formula III or other compounds of the present invention as described
above.

[0074] Preferably, the mammal is a human and the viral infection is a
flavivirus infection. More preferably, the flavivirus is selected from
the group consisting of Dengue virus, West Nile virus, yellow fever
virus, Japanese encephalitis virus, and tick-borne encephalitis virus.
Most preferably, the flavivirus is a Dengue virus selected from the group
consisting of DEN-1, DEN-2, DEN-3, and DEN-4.

[0075] Preferably, the viral infection is associated with a condition
selected from the group consisting of Dengue fever, Yellow fever, West
Nile, St. Louis encephalitis, Hepatitis C, Murray Valley encephalitis,
and Japanese encephalitis. Most preferably, the viral infection is
associated with Dengue fever wherein said Dengue fever is selected from
the group consisting of classical dengue fever and dengue hemorrhagic
fever.

[0076] The method of the present invention may also comprise
co-administration of: a) other antivirals; b) vaccines; and/or c)
interferons or pegylated interferons.

[0077] The present invention also provides for methods of synthesis of
compounds of the present invention, in particular
3-amino-6,7,8,9-tetrahydro-5H-1-thia-7,10-diaza-cyclohepta[f]indene-2-car-
boxylic acid (5-phenyl-[1,3,4]thiadiazol-2-yl)-amide and
3-amino-6,7,8,9-tetrahydro-5H-1-thia-6,10-diaza-cyclohepta[f]indene-2-car-
boxylic acid (5-phenyl-[1,3,4]thiadiazol-2-yl)-amide. These methods of
synthesis are provided below in Examples 14 and 15.

[0078] Novel Intermediates in the synthesis of the compounds of the
present invention include but are not limited to each of tert-butyl
(4E)-4-(hydroxymethylene)-5-oxoazepane-1-carboxylate; tert-butyl
(3E)-3-(hydroxymethylene)-4-oxoazepane-1-carboxylate; tert-butyl
3-cyano-2-thioxo-1,2,5,6,8,9-hexahydro-7H-pyrido[2,3-d]azepine-7-carboxyl-
ate; tert-butyl
3-cyano-2-thioxo-1,2,5,7,8,9-hexahydro-6H-pyrido[3,2-c]azepine-6-carboxyl-
ate; and 3-amino-7-tert-butyloxycarbonyl-6,7,8,9-tetrahydro-5H-1-thia-7,10-
-diaza-cyclohepta[f]indene-2-carboxylic acid
(5-phenyl-[1,3,4]thiadiazol-2-yl)-amide; and
3-amino-6-tert-butyloxycarbonyl-6,7,8,9-tetrahydro-5H-1-thia-6,10-diaza-c-
yclohepta[f]indene-2-carboxylic acid
(5-phenyl-[1,3,4]thiadiazol-2-yl)-amide.

DEFINITIONS

[0079] In accordance with this detailed description, the following
abbreviations and definitions apply. It must be noted that as used
herein, the singular forms "a", "an", and "the" include plural referents
unless the context clearly dictates otherwise.

[0080] The publications discussed herein are provided solely for their
disclosure. Nothing herein is to be construed as an admission regarding
antedating the publications. Further, the dates of publication provided
may be different from the actual publications dates, which may need to be
independently confirmed.

[0081] Where a range of values is provided, it is understood that each
intervening value is encompassed. The upper and lower limits of these
smaller ranges may independently be included in the smaller, subject to
any specifically-excluded limit in the stated range. Where the stated
range includes one or both of the limits, ranges excluding either both of
those included limits are also included in the invention. Also
contemplated are any values that fall within the cited ranges.

[0082] Unless defined otherwise, all technical and scientific terms used
herein have the same meaning as commonly understood by one of ordinary
skill in the art. Any methods and materials similar or equivalent to
those described herein can also be used in practice or testing. All
publications mentioned herein are incorporated herein by reference to
disclose and describe the methods and/or materials in connection with
which the publications are cited.

[0083] By "patient" or "subject" is meant to include any mammal. A
"mammal", for purposes of treatment, refers to any animal classified as a
mammal, including but not limited to, humans, experimental animals
including rats, mice, and guinea pigs, domestic and farm animals, and
zoo, sports, or pet animals, such as dogs, horses, cats, cows, and the
like.

[0084] The term "efficacy" as used herein refers to the effectiveness of a
particular treatment regime. Efficacy can be measured based on change of
the course of the disease in response to an agent.

[0085] The term "success" as used herein in the context of a chronic
treatment regime refers to the effectiveness of a particular treatment
regime. This includes a balance of efficacy, toxicity (e.g., side effects
and patient tolerance of a formulation or dosage unit), patient
compliance, and the like. For a chronic administration regime to be
considered "successful" it must balance different aspects of patient care
and efficacy to produce a favorable patient outcome.

[0086] The terms "treating", "treatment", and the like are used herein to
refer to obtaining a desired pharmacological and physiological effect.
The effect may be prophylactic in terms of preventing or partially
preventing a disease, symptom, or condition thereof and/or may be
therapeutic in terms of a partial or complete cure of a disease,
condition, symptom, or adverse effect attributed to the disease. The term
"treatment", as used herein, covers any treatment of a disease in a
mammal, such as a human, and includes: (a) preventing the disease from
occurring in a subject which may be predisposed to the disease but has
not yet been diagnosed as having it, i.e., causing the clinical symptoms
of the disease not to develop in a subject that may be predisposed to the
disease but does not yet experience or display symptoms of the disease;
(b) inhibiting the disease, i.e., arresting or reducing the development
of the disease or its clinical symptoms; and (c) relieving the disease,
i.e., causing regression of the disease and/or its symptoms or condition.
Treating a patient's suffering from disease related to a pathological
inflammation is contemplated. Preventing, inhibiting, or relieving
adverse effects attributed to pathological inflammation over long periods
of time and/or are such caused by the physiological responses to
inappropriate inflammation present in a biological system over long
periods of time are also contemplated.

[0089] "Alkenyl" refers to alkenyl group preferably having from 2 to 10
carbon atoms and more preferably 2 to 6 carbon atoms and having at least
1 and preferably from 1-2 sites of alkenyl unsaturation.

[0090] "Alkoxy" refers to the group "alkyl-O--" which includes, by way of
example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy,
sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.

[0091] "Alkyl" refers to linear or branched alkyl groups having from 1 to
10 carbon atoms, alternatively 1 to 6 carbon atoms. This term is
exemplified by groups such as methyl, t-butyl, n-heptyl, octyl and the
like.

[0092] "Amino" refers to the group --NH2.

[0093] "Aryl" or "Ar" refers to an unsaturated aromatic carbocyclic group
of from 6 to 14 carbon atoms having a single ring (e.g., phenyl) or
multiple condensed rings (e.g., naphthyl or anthryl) which condensed
rings may or may not be aromatic (e.g., 2-benzoxazolinone,
2H-1,4-benzoxazin-3(4H)-one, and the like) provided that the point of
attachment is through an aromatic ring atom.

[0095] "Cycloalkyl" refers to cyclic alkyl groups of from 3 to 8 carbon
atoms having a single cyclic ring including, by way of example,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl and the
like. Excluded from this definition are multi-ring alkyl groups such as
adamantanyl, etc.

[0096] "Halo" or "halogen" refers to fluoro, chloro, bromo and iodo.

[0097] "Heteroaryl" refers to an aromatic carbocyclic group of from 2 to
10 carbon atoms and 1 to 4 heteroatoms selected from the group consisting
of oxygen, nitrogen and sulfur within the ring or oxides thereof. Such
heteroaryl groups can have a single ring (e.g., pyridyl or furyl) or
multiple condensed rings (e.g., indolizinyl or benzothienyl) wherein one
or more of the condensed rings may or may not be aromatic provided that
the point of attachment is through an aromatic ring atom. Additionally,
the heteroatoms of the heteroaryl group may be oxidized, i.e., to form
pyridine N-oxides or 1,1-dioxo-1,2,5-thiadiazoles and the like.
Additionally, the carbon atoms of the ring may be substituted with an oxo
(═O). The term "heteroaryl having two nitrogen atoms in the
heteroaryl, ring" refers to a heteroaryl group having two, and only two,
nitrogen atoms in the heteroaryl ring and optionally containing 1 or 2
other heteroatoms in the heteroaryl ring, such as oxygen or sulfur.

[0100] "Optionally substituted" means that the recited group may be
unsubstituted or the recited group may be substituted.

[0101] "Pharmaceutically-acceptable carrier" means a carrier that is
useful in preparing a pharmaceutical composition or formulation that is
generally safe, non-toxic, and neither biologically nor otherwise
undesirable, and includes a carrier that is acceptable for veterinary use
as well as human pharmaceutical use.

[0102] "Pharmaceutically-acceptable cation" refers to the cation of a
pharmaceutically-acceptable salt.

[0103] "Pharmaceutically-acceptable salt" refers to salts which retain the
biological effectiveness and properties of compounds which are not
biologically or otherwise undesirable. Pharmaceutically-acceptable salts
refer to pharmaceutically-acceptable salts of the compounds, which salts
are derived from a variety of organic and inorganic counter ions well
known in the art and include, by way of example only, sodium, potassium,
calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when
the molecule contains a basic functionality, salts of organic or
inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate,
acetate, maleate, oxalate and the like.

[0107] A compound may act as a pro-drug. Pro-drug means any compound which
releases an active parent drug in vivo when such pro-drug is administered
to a mammalian subject. Pro-drugs are prepared by modifying functional
groups present in such a way that the modifications may be cleaved in
vivo to release the parent compound. Pro-drugs include compounds wherein
a hydroxy, amino, or sulfhydryl group is bonded to any group that may be
cleaved in vivo to regenerate the free hydroxyl, amino, or sulfhydryl
group, respectively. Examples of pro-drugs include, but are not limited
to esters (e.g., acetate, formate, and benzoate derivatives), carbamates
(e.g., N,N-dimethylamino-carbonyl) of hydroxy functional groups, and the
like.

[0108] "Treating" or "treatment" of a disease includes:

(1) preventing the disease, i.e. causing the clinical symptoms of the
disease not to develop in a mammal that may be exposed to or predisposed
to the disease but does not yet experience or display symptoms of the
disease, (2) inhibiting the disease, i.e., arresting or reducing the
development of the disease or its clinical symptoms, or (3) relieving the
disease, i.e., causing regression of the disease or its clinical
symptoms.

[0109] A "therapeutically-effective amount" means the amount of a compound
that, when administered to a mammal for treating a disease, is sufficient
to effect such treatment for the disease. The "therapeutically-effective
amount" will vary depending on the compound, the disease, and its
severity and the age, weight, etc., of the mammal to be treated.

Pharmaceutical Formulations of the Compounds

[0110] "Pharmaceutical composition" refers to a composition intended and
suitable for human or animal administration. A composition containing a
compound of the present invention dissolved in a solvent such as water,
organic solvent, alcohol or DMSO for the intended purpose of in-vitro
testing or for any type of testing outside of an animal or human body is
not considered a pharmaceutical composition as defined herein.

[0111] In general, compounds will be administered in a
therapeutically-effective amount by any of the accepted modes of
administration for these compounds. The compounds can be administered by
a variety of routes, including, but not limited to, oral, parenteral
(e.g., subcutaneous, subdural, intravenous, intramuscular, intrathecal,
intraperitoneal, intracerebral, intraarterial, or intralesional routes of
administration), topical, intranasal, localized (e.g., surgical
application or surgical suppository), rectal, and pulmonary (e.g.,
aerosols, inhalation, or powder). Accordingly, these compounds are
effective as both injectable and oral compositions. The compounds can be
administered continuously by infusion or by bolus injection.

[0112] The actual amount of the compound, i.e., the active ingredient,
will depend on a number of factors, such as the severity of the disease,
i.e., the condition or disease to be treated, age, and relative health of
the subject, the potency of the compound used, the route and form of
administration, and other factors.

[0113] Toxicity and therapeutic efficacy of such compounds can be
determined by standard pharmaceutical procedures in cell cultures or
experimental animals, e.g., for determining the LD50 (the dose
lethal to 50% of the population) and the ED50 (the dose
therapeutically effective in 50% of the population). The dose ratio
between toxic and therapeutic effects is the therapeutic index and it can
be expressed as the ratio LD50/ED50.

[0114] The data obtained from the cell culture assays and animal studies
can be used in formulating a range of dosage for use in humans. The
dosage of such compounds lies within a range of circulating
concentrations that include the ED50 with little or no toxicity. The
dosage may vary within this range depending upon the dosage form employed
and the route of administration utilized. For any compound used, the
therapeutically-effective dose can be estimated initially from cell
culture assays. A dose may be formulated in animal models to achieve a
circulating plasma concentration range which includes the IC50
(i.e., the concentration of the test compound which achieves a
half-maximal inhibition of symptoms) as determined in cell culture. Such
information can be used to more accurately determine useful doses in
humans. Levels in plasma may be measured, for example, by high
performance liquid chromatography.

[0115] The amount of the pharmaceutical composition administered to the
patient will vary depending upon what is being administered, the purpose
of the administration, such as prophylaxis or therapy, the state of the
patient, the manner of administration, and the like. In therapeutic
applications, compositions are administered to a patient already
suffering from a disease in an amount sufficient to cure or at least
partially arrest the symptoms of the disease and its complications. An
amount adequate to accomplish this is defined as
"therapeutically-effective dose." Amounts effective for this use will
depend on the disease condition being treated as well as by the judgment
of the attending clinician depending upon factors such as the severity of
the inflammation, the age, weight, and general condition of the patient,
and the like.

[0116] The compositions administered to a patient are in the form of
pharmaceutical compositions described supra. These compositions may be
sterilized by conventional sterilization techniques, or may be sterile
filtered. The resulting aqueous solutions may be packaged for use as is,
or lyophilized, the lyophilized preparation being combined with a sterile
aqueous carrier prior to administration. It will be understood that use
of certain of the foregoing excipients, carriers, or stabilizers will
result in the formation of pharmaceutical salts.

[0117] The active compound is effective over a wide dosage range and is
generally administered in a pharmaceutically- or
therapeutically-effective amount. The therapeutic dosage of the compounds
will vary according to, for example, the particular use for which the
treatment is made, the manner of administration of the compound, the
health and condition of the patient, and the judgment of the prescribing
physician. For example, for intravenous administration, the dose will
typically be in the range of about 0.5 mg to about 100 mg per kilogram
body weight. Effective doses can be extrapolated from dose-response
curves derived from in vitro or animal model test systems. Typically, the
clinician will administer the compound until a dosage is reached that
achieves the desired effect.

[0118] When employed as pharmaceuticals, the compounds are usually
administered in the form of pharmaceutical compositions. Pharmaceutical
compositions contain as the active ingredient one or more of the
compounds above, associated with one or more pharmaceutically-acceptable
carriers or excipients. The excipient employed is typically one suitable
for administration to human subjects or other mammals. In making the
compositions, the active ingredient is usually mixed with an excipient,
diluted by an excipient, or enclosed within a carrier which can be in the
form of a capsule, sachet, paper or other container. When the excipient
serves as a diluent, it can be a solid, semi-solid, or liquid material,
which acts as a vehicle, carrier, or medium for the active ingredient.
Thus, the compositions can be in the form of tablets, pills, powders,
lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions,
syrups, aerosols (as a solid or in a liquid medium), ointments
containing, for example, up to 10% by weight of the active compound, soft
and hard gelatin capsules, suppositories, sterile injectable solutions,
and sterile packaged powders.

[0119] In preparing a formulation, it may be necessary to mill the active
compound to provide the appropriate particle size prior to combining with
the other ingredients. If the active compound is substantially insoluble,
it ordinarily is milled to a particle size of less than 200 mesh. If the
active compound is substantially water soluble, the particle size is
normally adjusted by milling to provide a substantially uniform
distribution in the formulation, e.g., about 40 mesh.

[0120] Some examples of suitable excipients include lactose, dextrose,
sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate,
alginates, tragacanth, gelatin, calcium silicate, microcrystalline
cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and
methyl cellulose. The formulations can additionally include: lubricating
agents such as talc, magnesium stearate, and mineral oil; wetting agents;
emulsifying and suspending agents; preserving agents such as methyl- and
propylhydroxy-benzoates; sweetening agents; and flavoring agents. The
compositions of the invention can be formulated so as to provide quick,
sustained, or delayed-release of the active ingredient after
administration to the patient by employing procedures known in the art.

[0121] The quantity of active compound in the pharmaceutical composition
and unit dosage form thereof may be varied or adjusted widely depending
upon the particular application, the manner or introduction, the potency
of the particular compound, and the desired concentration. The term "unit
dosage forms" refers to physically-discrete units suitable as unitary
dosages for human subjects and other mammals, each unit containing a
predetermined quantity of active material calculated to produce the
desired therapeutic effect, in association with a suitable pharmaceutical
excipient.

[0122] The compound can be formulated for parenteral administration in a
suitable inert carrier, such as a sterile physiological saline solution.
The dose administered will be determined by route of administration.

[0123] Administration of therapeutic agents by intravenous formulation is
well known in the pharmaceutical industry. An intravenous formulation
should possess certain qualities aside from being just a composition in
which the therapeutic agent is soluble. For example, the formulation
should promote the overall stability of the active ingredient(s), also,
the manufacture of the formulation should be cost-effective. All of these
factors ultimately determine the overall success and usefulness of an
intravenous formulation.

[0125] The presence of a buffer is necessary to maintain the aqueous pH in
the range of from about 4 to about 8. The buffer system is generally a
mixture of a weak acid and a soluble salt thereof, e.g., sodium
citrate/citric acid; or the monocation or dication salt of a dibasic
acid, e.g., potassium hydrogen tartrate; sodium hydrogen tartrate,
phosphoric acid/potassium dihydrogen phosphate, and phosphoric
acid/disodium hydrogen phosphate.

[0126] The amount of buffer system used is dependent on (1) the desired
pH; and (2) the amount of drug. Generally, the amount of buffer used is
able to maintain a formulation pH in the range of 4 to 8. Generally, a
1:1 to 10:1 mole ratio of buffer (where the moles of buffer are taken as
the combined moles of the buffer ingredients, e.g., sodium citrate and
citric acid) to drug is used.

[0127] A useful buffer is sodium citrate/citric acid in the range of 5 to
50 mg per ml. sodium citrate to 1 to 15 mg per ml. citric acid,
sufficient to maintain an aqueous pH of 4-6 of the composition.

[0128] The buffer agent may also be present to prevent the precipitation
of the drug through soluble metal complex formation with dissolved metal
ions, e.g., Ca, Mg, Fe, Al, Ba, which may leach out of glass containers
or rubber stoppers or be present in ordinary tap water. The agent may act
as a competitive complexing agent with the drug and produce a soluble
metal complex leading to the presence of undesirable particulates.

[0129] In addition, the presence of an agent, e.g., sodium chloride in an
amount of about of 1-8 mg/ml, to adjust the tonicity to the same value of
human blood may be required to avoid the swelling or shrinkage of
erythrocytes upon administration of the intravenous formulation leading
to undesirable side effects such as nausea or diarrhea and possibly to
associated blood disorders. In general, the tonicity of the formulation
matches that of human blood which is in the range of 282 to 288 mOsm/kg,
and in general is 285 mOsm/kg, which is equivalent to the osmotic
pressure corresponding to a 0.9% solution of sodium chloride.

[0130] An intravenous formulation can be administered by direct
intravenous injection, i.v. bolus, or can be administered by infusion by
addition to an appropriate infusion solution such as 0.9% sodium chloride
injection or other compatible infusion solution.

[0131] The compositions are preferably formulated in a unit dosage form,
each dosage containing from about 5 to about 100 mg, more usually about
10 to about 30 mg, of the active ingredient. The term "unit dosage forms"
refers to physically discrete units suitable as unitary dosages for human
subjects and other mammals, each unit containing a predetermined quantity
of active material calculated to produce the desired therapeutic effect,
in association with a suitable pharmaceutical excipient.

[0132] The active compound is effective over a wide dosage range and is
generally administered in a pharmaceutically effective amount. It will be
understood, however, that the amount of the compound actually
administered will be determined by a physician, in the light of the
relevant circumstances, including the condition to be treated, the chosen
route of administration, the actual compound administered, the age,
weight, and response of the individual patient, the severity of the
patient's symptoms, and the like.

[0133] For preparing solid compositions such as tablets, the principal
active ingredient is mixed with a pharmaceutical excipient to form a
solid preformulation composition containing a homogeneous mixture of a
compound of the present invention. When referring to these preformulation
compositions as homogeneous, it is meant that the active ingredient is
dispersed evenly throughout the composition so that the composition may
be readily subdivided into equally effective unit dosage forms such as
tablets, pills and capsules. This solid preformulation is then subdivided
into unit dosage forms of the type described above containing from, for
example, 0.1 to about 2000 mg of the active ingredient.

[0134] The tablets or pills may be coated or otherwise compounded to
provide a dosage form affording the advantage of prolonged action. For
example, the tablet or pill can comprise an inner dosage and an outer
dosage component, the latter being in the form of an envelope over the
former. The two components can be separated by an enteric layer which
serves to resist disintegration in the stomach and permit the inner
component to pass intact into the duodenum or to be delayed in release. A
variety of materials can be used for such enteric layers or coatings,
such materials including a number of polymeric acids and mixtures of
polymeric acids with such materials as shellac, cetyl alcohol, and
cellulose acetate.

[0135] The liquid forms in which the novel compositions may be
incorporated for administration orally or by injection include aqueous
solutions, suitably flavored syrups, aqueous or oil suspensions, and
flavored emulsions with edible oils such as cottonseed oil, sesame oil,
coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical
vehicles.

[0136] Compositions for inhalation or insufflation include solutions and
suspensions in pharmaceutically-acceptable, aqueous or organic solvents,
or mixtures thereof, and powders. The liquid or solid compositions may
contain suitable pharmaceutically-acceptable excipients as described
supra. Compositions in pharmaceutically-acceptable solvents may be
nebulized by use of inert gases. Nebulized solutions may be breathed
directly from the nebulizing device or the nebulizing device may be
attached to a face masks tent, or intermittent positive pressure
breathing machine. Solution, suspension, or powder compositions may be
administered from devices which deliver the formulation in an appropriate
manner.

[0138] The compounds can be administered in a sustained-release form, for
example a depot injection, implant preparation, or osmotic pump, which
can be formulated in such a manner as to permit a sustained-release of
the active ingredient. Implants for sustained-release formulations are
well-known in the art. Implants may be formulated as, including but not
limited to, microspheres, slabs, with biodegradable or non-biodegradable
polymers. For example, polymers of lactic acid and/or glycolic acid form
an erodible polymer that is well-tolerated by the host.

[0139] Transdermal delivery devices ("patches") may also be employed. Such
transdermal patches may be used to provide continuous or discontinuous
infusion of the compounds in controlled amounts. The construction and use
of transdermal patches for the delivery of pharmaceutical agents is well
known in the art. See, e.g., U.S. Pat. No. 5,023,252, issued Jun. 11,
1991, herein incorporated by reference. Such patches may be constructed
for continuous, pulsatile, or on-demand delivery of pharmaceutical
agents.

[0140] Direct or indirect placement techniques may be used when it is
desirable or necessary to introduce the pharmaceutical composition to the
brain. Direct techniques usually involve placement of a drug delivery
catheter into the host's ventricular system to bypass the blood-brain
barrier. One such implantable delivery system used for the transport of
biological factors to specific anatomical regions of the body is
described in U.S. Pat. No. 5,011,472, which is herein incorporated by
reference.

[0141] Indirect techniques usually involve formulating the compositions to
provide for drug latentiation by the conversion of hydrophilic drugs into
lipid-soluble drugs. Latentiation is generally achieved through blocking
of the hydroxy, carbonyl, sulfate, and primary amine groups present on
the drug to render the drug more lipid-soluble and amenable to
transportation across the blood-brain barrier. Alternatively, the
delivery of hydrophilic drugs may be enhanced by intra-arterial infusion
of hypertonic solutions which can transiently open the blood-brain
barrier.

[0142] In order to enhance serum half-life, the compounds may be
encapsulated, introduced into the lumen of liposomes, prepared as a
colloid, or other conventional techniques may be employed which provide
an extended serum half-life of the compounds. A variety of methods are
available for preparing liposomes, as described in, e.g., Szoka et al.,
U.S. Pat. Nos. 4,235,871, 4,501,728 and 4,837,028 each of which is
incorporated herein by reference.

[0143] Pharmaceutical compositions are suitable for use in a variety of
drug delivery systems. Suitable formulations for use in the present
invention are found in Remington's Pharmaceutical Sciences, Mace
Publishing Company, Philadelphia, Pa., 17th ed. (1985).

[0144] In the examples below, if an abbreviation is not defined above, it
has its generally accepted meaning. Further, all temperatures are in
degrees Celsius (unless otherwise indicated). The following Methods were
used to prepare the compounds set forth below as indicated.

Example 1

Formulation 1

[0145] Hard gelatin capsules containing the following ingredients are
prepared:

[0152] The active ingredient, starch, and cellulose are passed through a
No. 20 mesh U.S. sieve and mixed thoroughly. The solution of
polyvinyl-pyrrolidone is mixed with the resultant powders, which are then
passed through a 16 mesh U.S. sieve. The granules so produced are dried
at 50° to 60° C. and passed through a 16 mesh U.S. sieve.
The sodium carboxymethyl starch, magnesium stearate, and talc, previously
passed through a No. 30 mesh U.S. sieve, are then added to the granules,
which after mixing, are compressed on a tablet machine to yield tablets
each weighing 150 mg.

Example 5

Formulation 5

[0153] Capsules, each containing 40 mg of medicament, are made as follows:

[0156] The active ingredient is passed through a No. 60 mesh U.S. sieve
and suspended in the saturated fatty acid glycerides previously melted
using the minimum heat necessary. The mixture is then poured into a
suppository mold of nominal 2.0 g capacity and allowed to cool.

Example 7

Formulation 7

[0157] Suspensions, each containing 50 mg of medicament per 5.0 ml dose,
are made as follows:

[0158] The medicament, sucrose, and xanthan gum are blended, passed
through a NO. 10 mesh U.S. sieve, and then mixed with a previously made
solution of the microcrystalline cellulose and sodium carboxymethyl
cellulose in water. The sodium benzoate, flavor, and color are diluted
with some of the water and added with stirring. Sufficient water is then
added to produce the required volume.

Example 8

Formulation 8

[0159] Hard gelatin tablets, each containing 15 mg of active ingredient,
are made as follows:

[0162] Therapeutic compound compositions generally are placed into a
container having a sterile access port, for example, an intravenous
solution bag or vial having a stopper pierceable by a hypodermic
injection needle or similar sharp instrument.

[0164] The white soft paraffin is heated until molten. The liquid paraffin
and emulsifying wax are incorporated and stirred until dissolved. The
active ingredient is added and stirring is continued until dispersed. The
mixture is then cooled until solid.

Example 11

Formulation 11

[0165] An aerosol formulation may be prepared as follows: A solution of
the candidate compound in 0.5% sodium bicarbonate/saline (w/v) at a
concentration of 30.0 mg/mL is prepared using the following procedure:

Development of a High-Throughput Screening Assay for Measurement of Dengue
Virus-Induced Cytopathic Effect

[0170] A sensitive and reproducible high-throughput screening (HTS) assay
has been established to measure dengue virus-induced cytopathic effect
(CPE). To determine the amount of dengue virus stock required to produce
complete CPE in 5 days, Vero cell monolayers were seeded on 96-well
plates and infected with 10-fold serial dilutions of the dengue virus
stock representing a multiplicity of infection (MOI) of approximately
0.001 PFU/cell to 0.1 PFU/cell. At 5 days post-infection, the cultures
were fixed with 5% glutaraldehyde and stained with 0.1% crystal violet.
Virus-induced CPE was quantified spectrophotometrically at OD570.
From this analysis, an MOI of 0.1 PFU/cell of dengue virus stock was
chosen for use in the HTS assay. To establish the signal-to-noise ratio
(S/N) of the 96-well assay and evaluate the well-to-well and
assay-to-assay variability, five independent experiments were performed.
Vero cell monolayers were infected with 0.1 PFU/cell of dengue virus
stock. Each plate contained the following controls: quadruplicate
virus-infected wells, quadruplicate uninfected cell wells and a dose
response curve in duplicate for ribavirin at 500, 250, 125 and 62 μM,
as reference standards. At day 5 post-infection, the plates were
processed as described above.

[0171] The dengue virus CPE assay was used to evaluate compounds from the
SIGA chemical library for those that inhibit dengue virus-induced CPE.
Each evaluation run consisted of 48 96-well plates with 80 compounds per
plate to generate 4,608 data points per run. At this throughput we are
capable of evaluating 200,000 compounds in about 52 weeks. Compounds were
dissolved in DMSO and diluted in medium such that the final concentration
in each well was 5 μM compound and 0.5% DMSO. The compounds were added
robotically to the culture medium using the PerkinElmer MultiPROBE®
II HT PLUS robotic system. Following compound addition, cultures were
infected with dengue virus (DEN-2 strain New Guinea C). After 5 days
incubation, plates were processed and CPE quantified on a PerkinElmer
EnVision II plate reader system.

[0172] The results of these experiments indicated that the 96-well assay
format is robust and reproducible. The S/N ratio (ratio of signal of cell
control wells (signal) to virus control wells (noise)) was 5.0±1.2.
The well-to-well variability was determined for each individual plate and
found to have a coefficient of variance of less than 10% for both
positive control and negative control wells, and overall assay-to-assay
variability was less than 15%. Using this assay, the EC50 values for
ribavirin were determined to be 125±25 μM, respectively. The
effectiveness of ribavirin against dengue varies with the cell type used,
but the values we obtained were within the range of published values for
this compound (2, 13, 28). Taken together, these results show that a
sensitive and reproducible HTS assay has been successfully developed to
evaluate our compound library for inhibitors of dengue virus replication.

Example 13

Determining Anti-Dengue-2 Activity of Compounds of the Invention

[0173] The assay described in Example 12 was the basis of a
high-throughput screen for dengue virus inhibitors, against which a
library of 210,000 compounds was tested. Compounds that inhibited dengue
virus induced CPE by at least 50% were further investigated for chemical
tractability, potency, and selectivity.

[0174] Initially, the chemical structures of the hit compounds were
examined for chemical tractability. A chemically tractable compound is
defined as one that is synthetically accessible using reasonable chemical
methodology, and which possesses chemically stable functionalities and
potential drug-like qualities. Hits that passed this medicinal chemistry
filter were evaluated for their potency. Compound potency was determined
by evaluating inhibitory activity across a broad range of concentrations.
Nonlinear regression was used to generate best-fit inhibition curves and
to calculate the 50% effective concentration (EC50). The selectivity
or specificity of a given compound is typically expressed as a ratio of
its cytotoxicity to its biological effect. A cell proliferation assay is
used to calculate a 50% cytotoxicity concentration (CC50); the ratio
of this value to the EC50 is referred to as the therapeutic index
(T.I.=CC50/EC50). Two types of assays have been used to
determine cytotoxicity, both of which are standard methods for
quantitating the reductase activity produced in metabolically active
cells (22). One is a colorimetric method that measures the reduction of
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), and
the other uses fluorimetry to measure the reduction of resazurin (Alamar
Blue). Selectivity could be further characterized by assessing the
inhibitory action against viruses from unrelated virus families. Sixteen
quality dengue hits were discovered in the pool of initial hits from the
HTS screening, all with EC50 values below 25 μM. Verification
that these compounds act against each of the four serotypes of dengue was
done with yield assays carried out at several drug concentrations, and
the titer determined for each.

[0175] Compounds that were active in the primary screen were tested for
activity in viral yield assays. Table 1 shows some of the compounds that
were tested for activity against Dengue-2 (Strain New Guinea C) in a
viral yield assay at a range of concentrations. Vero cells in 12-well
plates were infected with dengue-2 virus at a multiplicity of infection
(MOI) of 0.1, treated with compound (or DMSO as a control), incubated at
37° C., harvested 48 hours post infection and titered on Vero
cells as described above. The EC50 was calculated through ExcelFit.
Activities against other serotypes of dengue virus were determined in a
similar way.

[0176] Compound 1 was identified as one of the most potent and selective
compounds from within the pool of the initial quality hits, with activity
against all four serotypes of dengue. Chemical analogs of this compound
were obtained, and these analogs were tested as described in order to
define the relationship between chemical structure and biological
activity (see Table 1). All of the compounds in Table 1, labeled A or B,
are active against dengue with EC50 values at or below 25 μM.

TABLE-US-00015
TABLE 3
Novel Compounds of Formula III activity against
Dengue Virus in Vero cells.
Activity (EC50 in μM)
A: EC50 ≦ 5 μM; B: 5 < EC50 ≦ 25 μM;
C: EC50 > 25 μM; n.d.: not determined
Cmpd DENV-1 DENV-2 DENV-3 DENV-4
285 A A A A
289 A A A A
293 A A A A
294 A A A A
295 A A A A
296 A A A A
297 A A A A
298 A A A A
299 B B n.d. B
300 A A A A
302 A A B A
303 B A B A
304 A A A A
305 A A B A
307 A A A A
308 n.d. A n.d. n.d.
309 A A A A
310 A A A A
311 A A A A
312 A A A A
313 n.d. A n.d. n.d.
314 n.d. A n.d. n.d.
315 n.d. A n.d. n.d.
321 A A A A
358 A A B C
359 A A C B
360 C A C A
361 A A A C
362 B B C C
363 B A C C

TABLE-US-00017
TABLE 5
Activity against Dengue virus of novel compounds of
the present invention outside the scope of Formula III.
Activity (EC50 in μM)
A: EC50 ≦ 5 μM; B: 5 < EC50 ≦ 25 μM;
C: EC50 > 25 μM; n.d.: not determined
Cmpd DENV-1 DENV-2 DENV-3 DENV-4
281 n.d. B n.d. n.d.
282 n.d. B n.d. n.d.
283 n.d. A n.d. n.d.
284 A A B C
286 n.d. A n.d. n.d.
287 n.d. B n.d. n.d.
288 A A B A
290 n.d. A n.d. n.d.
291 n.d. B n.d. n.d.
292 A A A A
301 A A B A
306 A A A A
316 n.d. A n.d. n.d.
317 n.d. A n.d. n.d.
318 n.d. A n.d. n.d.
319 n.d. A n.d. n.d.
320 n.d. A n.d. n.d.
322 A A A A
323 n.d. A n.d. n.d.
324 n.d. A n.d. n.d.
325 A A A A
326 n.d. A n.d. n.d.
327 A A A A
328 A A B A
329 A A B A
330 B A B B
331 A A A B
332 A A A A
333 A A A A
334 n.d. A n.d. n.d.
335 A A A A
336 A A A A
337 A A A A
338 A A A A
339 A A A A
340 A A A A
341 A A A A
342 A A A A
343 A A A A
344 A A A A
345 A A A A
346 n.d. A n.d. n.d.
347 n.d. A n.d. n.d.
348 n.d. A n.d. n.d.
349 A A A A
350 A A A A
351 n.d. A n.d. n.d.
352 A A A A
353 A A A A
354 n.d. B n.d. n.d.
355 n.d. A n.d. n.d.
356 n.d. B n.d. n.d.
357 n.d. A n.d. n.d.

TABLE-US-00019
TABLE 7
Activity against Dengue virus of compounds of the present invention.
Activity (EC50 in μM)
A: EC50 ≦ 5 μM; B: 5 < EC50 ≦ 25 μM;
C: EC50 > 25 μM; n.d.: not determined
Cmpd DENV-1 DENV-2 DENV-3 DENV-4
364 A A A A
365 B A B C
366 n.d. A n.d. n.d.
367 n.d. A n.d. n.d.
368 A A A A
369 B A A A
370 n.d. A n.d. n.d.
371 n.d. A n.d. n.d.
372 n.d. A n.d. n.d.
373 A A A A
374 n.d. A n.d. n.d.
375 n.d. B n.d. n.d.
376 n.d. B n.d. n.d.
377 n.d. A n.d. n.d.
378 n.d. A n.d. n.d.
379 n.d. A n.d. n.d.
380 n.d. A n.d. n.d.
381 n.d. B n.d. n.d.
382 n.d. B n.d. n.d.
383 n.d. A n.d. n.d.
384 n.d. B n.d. n.d.
385 n.d. B n.d. n.d.
386 n.d. A n.d. n.d.
387 n.d. B n.d. n.d.
388 n.d. A n.d. n.d.
389 n.d. B n.d. n.d.
390 n.d. A n.d. n.d.
391 n.d. A n.d. n.d.
392 n.d. B n.d. n.d.
393 n.d. B n.d. n.d.
394 n.d. A n.d. n.d.
395 n.d. B n.d. n.d.
396 n.d. B n.d. n.d.
397 n.d. A n.d. n.d.
398 n.d. B n.d. n.d.
399 n.d. A n.d. n.d.
400 n.d. A n.d. n.d.
401 n.d. C n.d. n.d.
402 n.d. C n.d. n.d.
403 n.d. A n.d. n.d.
404 n.d. A n.d. n.d.
405 n.d. A n.d. n.d.
406 n.d. A n.d. n.d.
407 n.d. C n.d. n.d.
408 n.d. C n.d. n.d.
409 n.d. C n.d. n.d.
410 n.d. A n.d. n.d.
411 A A A A
412 n.d. B n.d. n.d.
413 A A A A
414 A A A A
415 A A A A
416 n.d. A n.d. n.d.
417 A A A A
418 n.d. B n.d. n.d.
419 n.d. A n.d. n.d.
420 n.d. A n.d. n.d.
421 n.d. B n.d. n.d.
422 n.d. A n.d. n.d.
423 n.d. A n.d. n.d.
424 n.d. B n.d. n.d.
425 A A A A
426 n.d. A n.d. n.d.
427 A A A A
428 A A A A
429 A A A A
430 A A A A
431 A A A A
432 n.d. B n.d. n.d.
433 A A A A
434 A A A A
435 n.d. A n.d. n.d.
436 n.d. A n.d. n.d.
437 A A A A
438 A A A A
439 A A A A
440 n.d. B n.d. n.d.
441 A A A A
442 n.d. A n.d. n.d.
443 n.d. A n.d. n.d.
444 n.d. A n.d. n.d.
445 A A A A
446 A A A A
447 A A A A
448 A A A A
449 A A A A
450 A A A A
451 n.d. A n.d. n.d.
452 A A A A
453 A A A A
454 A A A A
455 A A A B
456 n.d. A n.d. n.d.
457 n.d. B n.d. n.d.
458 A A A A
459 A A A A
460 n.d. A n.d. n.d.
461 A A A A
462 A A A A
463 n.d. A n.d. n.d.
464 A A A A
465 A A A A
466 n.d. B n.d. n.d.
467 n.d. A n.d. n.d.
468 A A A A
469 A A A A
470 A A A A
471 A A A A
472 A A A A
473 A A A A
474 n.d. A n.d. n.d.
475 A A A A
476 A A A A
477 n.d. A n.d. n.d.
478 n.d. B n.d. n.d.
479 n.d. A n.d. n.d.
480 n.d. A n.d. n.d.
481 n.d. B n.d. n.d.
482 A A A A
483 A A A A
484 n.d. A n.d. n.d.
485 A A A A
486 A A A A
487 n.d. A n.d. n.d.
488 A A A A
489 A A A A
490 A A B A
491 C A B A
492 A A A A
493 A A A A
494 A A B A
495 A A A A
496 n.d. A n.d. n.d.
497 A A A A
498 A A A A
499 n.d. A n.d. n.d.
500 n.d. A n.d. n.d.
501 n.d. A n.d. n.d.
502 n.d. A n.d. n.d.
503 n.d. A n.d. n.d.
504 n.d. A n.d. n.d.
505 n.d. A n.d. n.d.
506 A A A A
507 A A A A
508 n.d. A n.d. n.d.
509 n.d. A n.d. n.d.
510 A A A A
511 n.d. A n.d. n.d.
512 A A A A
513 n.d. A n.d. n.d.
514 A A A A
515 n.d. A n.d. n.d.
516 n.d. A n.d. n.d.
517 n.d. A n.d. n.d.
518 n.d. A n.d. n.d.
519 n.d. A n.d. n.d.
520 n.d. A n.d. n.d.
521 n.d. A n.d. n.d.
522 A A A A
523 n.d. A n.d. n.d.
524 n.d. A n.d. n.d.
525 n.d. A n.d. n.d.
526 n.d. A n.d. n.d.
527 n.d. A n.d. n.d.
528 n.d. A n.d. n.d.
529 A A A A
530 A A A A
531 n.d. A n.d. n.d.
532 A A A A
533 A A A A
534 A A A A
535 A A A A
536 n.d. A n.d. n.d.
537 n.d. A n.d. n.d.
538 n.d. A n.d. n.d.
539 n.d. A n.d. n.d.
540 n.d. A n.d. n.d.
541 n.d. A n.d. n.d.
542 A A A A
543 A A A A
544 n.d. A n.d. n.d.
545 n.d. A n.d. n.d.
546 A A A A
547 A A A A
548 n.d. A n.d. n.d.
549 n.d. A n.d. n.d.
550 A A A A
551 n.d. A n.d. n.d.
552 n.d. A n.d. n.d.
553 n.d. A n.d. n.d.
554 n.d. A n.d. n.d.
555 A A A A
556 n.d. A n.d. n.d.
557 n.d. A n.d. n.d.
558 n.d. A n.d. n.d.
559 n.d. A A A
560 n.d. A n.d. n.d.
561 A A A A
562 n.d. A n.d. n.d.
563 n.d. A n.d. n.d.
564 n.d. A n.d. n.d.
565 n.d. A n.d. n.d.
566 A A A A
567 n.d. A n.d. n.d.
568 n.d. A n.d. n.d.
569 A A B A
570 A A A A
571 A A A A
572 A A A A
573 n.d. A n.d. n.d.
574 A A A A
575 A A A A
576 A A A A
577 A A A A
578 n.d. A n.d. n.d.
579 n.d. A n.d. n.d.
580 n.d. A n.d. n.d.
581 n.d. A n.d. n.d.
582 n.d. A n.d. n.d.
583 A A A A
584 n.d. A n.d. A
585 n.d. A n.d. n.d.
586 n.d. A n.d. n.d.
587 n.d. A n.d. n.d.
588 n.d. A n.d. n.d.
589 n.d. A n.d. n.d.
590 n.d. A n.d. n.d.
591 A A A A
592 n.d. A n.d. n.d.
593 n.d. A n.d. n.d.
594 n.d. A n.d. n.d.
595 n.d. A n.d. n.d.
596 A A A A
597 A A A A
598 A A A A
599 A A A A
600 A A A A
601 n.d. A n.d. n.d.
602 A A A B
603 n.d. A n.d. A
604 n.d. A n.d. n.d.
605 n.d. A n.d. n.d.

606 n.d. A n.d. n.d.
607 n.d. A n.d. n.d.
608 n.d. A n.d. n.d.
609 A A B B
610 n.d. A n.d. n.d.
611 n.d. A n.d. n.d.
612 A A A A
613 n.d. A n.d. n.d.
614 n.d. A n.d. n.d.
615 A A A A
616 A A A A
617 A A A A
618 A A n.d. n.d.
619 n.d. A n.d. n.d.
620 A A A A
621 n.d. A n.d. n.d.
622 n.d. A n.d. n.d.
623 n.d. A n.d. n.d.
624 n.d. A n.d. n.d.
625 A A A C
626 n.d. A n.d. n.d.
627 n.d. A n.d. n.d.
628 A A A A
629 n.d. A n.d. n.d.
630 A A A A
631 A A A A
632 n.d. A n.d. n.d.
633 n.d. A n.d. n.d.
634 n.d. A n.d. n.d.
635 A A C A
636 A A A A
637 n.d. A n.d. n.d.
638 A A A A
639 n.d. A n.d. n.d.
640 n.d. A n.d. n.d.
641 n.d. A n.d. n.d.
642 n.d. A n.d. n.d.
643 n.d. A n.d. n.d.
644 n.d. A n.d. n.d.
645 A A A A
646 A A A A
647 n.d. A n.d. n.d.
648 A A A A
649 n.d. A n.d. n.d.
650 A A A A
651 A A A A
652 A A A A
653 n.d. A n.d. n.d.
654 n.d. A n.d. n.d.
655 n.d. A n.d. n.d.

[0178] To a mixture of 5-phenyl-1,3,4-thiadiazol-2-amine (C1, 1.06 g, 6
mmol) and K2CO3 (0.83 g, 6 mmol) in anhydrous DMF (20 mL), was
added chloroacetyl chloride (C2, 0.48 mL, 6 mmol). The mixture was
stirred at room temperature for 4 h. The reaction mixture was then poured
into ice-water (100 mL), stirred, and then filtered. The resulting solid
was washed with water, and then dried in the oven under vacuum to afford
compound C3 (1.15 g, 76%) as a white solid.

[0179] A solution of tert-butyl 4-oxoazepane-1-carboxylate (C4, 2.56 g,
12.0 mmol) and N-[tert-butoxy(dimethylamino)methyl]-N,N-dimethylamine
(C5, 2.97 mL, 14.4 mmol) in THF (30 mL) was refluxed for 8 h. After
cooling, the reaction mixture was treated with water (20 mL), stirred at
room temperature for 15 min, and then extracted with EtOAc. The organic
layer was dried over Na2SO4, and concentrated under reduced
pressure to give C6 (major) and C7 (minor) as a colorless oil (2.63 g,
91%), which was used as a mixture in the next step reaction directly.

[0181] A mixture of C9 (750 mg, 2.46 mmol), C3 (623 mg, 2.46 mmol) and
sodium acetate (302 mg, 3.68 mmol) in EtOH (20 mL) was refluxed for 4 h.
After cooling, the reaction mixture was poured into water (100 mL),
stirred, and then filtered. The given solid was dried in the oven under
vacuum, and then recrystallized in EtOAc to afford compound C11 (500 mg,
39%) as a yellow solid. MS: MNa.sup.+=545.

[0186] A solution of 1-1 (19.04 g, 169.7 mmol) in anhydrous THF (50 mL)
was cooled to 0° C. A solution of LHMDS (1.0 M in THF, 190 mL, 190
mmol) was added dropwise, followed by ethyl formate (13.8 g, 186.3 mmol).
The resulting mixture was stirred for 3 h at 0° C. under N2
and quenched by slow addition of water (300 mL) and hexanes (200 mL). The
layers were separated, the aqueous layer was neutralized with 5% citric
acid (350 mL), followed by extraction with ethyl acetate (300
mL×2). Organic layers were combined, washed with water (300 mL),
brine (300 mL) and dried (Na2SO4). The solvent was removed
under reduced pressure and 1-2 was obtained as an oil (20.0 g, 84%
yield). This was used in the next step without further purification.

[0187] A mixture of 1-2 (18.0 g, 128.6 mmol), 2-cyanothioacetamide (12.9
g, 128.6 mmol) and a piperidine solution (122 mL, prepared from
piperidine (90 mL) and AcOH (53 mL) in water (125 mL)) in water (643 mL)
was heated to reflux for 15 minutes. Additional AcOH (193 mL) was added
and the reaction mixture was allowed to cool to room temperature slowly,
when compound 1-3 precipitated out as a red solid. The reaction mixture
was filtered and the cake was washed with water (100 mL) and dried under
vacuum (18.5 g, 70% yield).

General Procedure for the Preparation of 2-Bromoacetoamide

[0188] To a solution of the corresponding primary amine (25 mmol) in
anhydrous DCM (100 mL) was added a mixture of 2-bromoacetyl bromide (25
mmol) and triethylamine (30 mmol) in anhydrous DCM (20 mL) at -30°
C. under N2. After the addition, the reaction mixture was stirred at
room temperature for 1.5 h and then concentrated. The residue was
re-dissolved in acetone (50 mL), precipitated triethylamine hydrobromide
was removed by filtration, and the filtrate was evaporated to yield the
product. The product was further purified by trituration with diethyl
ether.

General Procedure for the Preparation of Final Products

[0189] To a slurry of compound 1-3 (1 mmol, 204 mg) in anhydrous EtOH (5
mL) was added the corresponding 2-bromoacetamide (1 mmol), followed by a
solution of sodium ethoxide in EtOH (2.6 M solution, 1.5 mmol, 0.58 mL)
at room temperature under N2. The reaction was heated to reflux for
2 hours and during that time, the desired product precipitated out. The
mixture was cooled to room temperature and filtered. The solid was washed
by EtOH (2 mL), diethyl ether (5 mL) and dried under vacuum to yield the
final products.

Example 17

Synthesis of Compounds 284, 286, 287 and 288

##STR00672##

[0191] To a slurry of 1-5 (100 mg, 0.333 mmol) in anhydrous EtOH (2.5 mL)
was added the corresponding sulfanylpyridine carbonitrile (1-7) followed
by a solution of sodium ethoxide in EtOH (2.6 M solution, 0.2 mL, 0.56
mmol) at room temperature under N2. The reaction was heated to
reflux for 2 hours and during that time, the desired product precipitated
out. The mixture was cooled to room temperature and filtered. The solid
was washed with EtOH (2 mL) and ether (5 mL), and dried under vacuum to
give the final compounds.

[0194] To a slurry of 1-3 (2.04 g, 10 mmol) in anhydrous EtOH (100 mL) was
added 1-5 (2.99 g, 10 mmol), followed by a solution of sodium ethoxide in
EtOH (2.6 M solution, 5.8 mL, 15 mmol) at room temperature under N2.
The reaction was heated to reflux for 2 hours and during that time, the
desired product precipitated out. The mixture was cooled to room
temperature and filtered. The solid was washed with EtOH (20 mL), diethyl
ether (50 mL), and dried under vacuum to give 1-6 (3.30 g, yield 78%).

[0196] To a solution of 1-6 (200 mg, 0.475 mmol) in anhydrous NMP (2 mL)
was added n-BuI (131 mg, 0.713 mmol) and the mixture was stirred at room
temperature for 1 h under N2. Then, DCM (100 mL) was added and the
mixture was washed with water (10 mL), aqueous saturated NaHCO3 (10
mL), brine (10 mL) and dried (Na2SO4). Most of the solvent was
removed under reduced pressure and the precipitated solid was filtered.
The cake was washed with diethyl ether (10 mL) and dried under vacuum to
yield 289 (70 mg, 31% yield).

[0198] To a solution of intermediate 1-6 (0.42 g, 1 mmol) and
triethylamine (2 mL) in N-methylpyrrolidinone (20 mL) was added
N(Boc)-2-bromoethylamine (1.8 g, 8.0 mmol) and the contents were heated
at 100° C. for 16 h. The reaction mixture was cooled to room
temperature and poured into ice-cold water. The solid obtained was
filtered and air-dried to give the free base (0.23 g). Treatment of the
free base with 2M HCl in diethyl ether (10 mL) at room temperature
overnight followed by filtration afforded 294 in the HCl salt form (0.19
g, 38% overall yield).

[0199] To a solution of intermediate 1-6 (0.63 g, 1.5 mmol) and TEA (1 mL)
in anhydrous DCM (30 mL) at 0° C. was added methylmalonyl chloride
(0.4 g, 3.0 mmol, 2.0 eq) dropwise and the contents were slowly warmed to
room temperature and stirred for 24 h. The organic portion was washed
with 1M NaOH, brine, dried (Na2SO4), filtered and concentrated.
The crude methyl ester was stirred with 1M LiOH (4 mL) in THF (12 mL) and
water (4 mL) at room temperature overnight. Most of the THF was removed
under vacuum and the solid obtained was filtered, dried and treated with
sodium methoxide (1.0 eq) in MTBE at room temperature overnight. The
solid obtained was filtered and dried under vacuum to give the sodium
salt of 295 (0.3 g, 38% overall yield) as a brown solid.

[0202] To a solution of 1-6 (200 mg, 0.475 mmol) in anhydrous NMP (2 mL)
was added CH3I (102 mg, 0.712 mmol) and stirred for 1 hour at room
temperature under N2. Then, DCM (100 mL) was added and the mixture
was washed with water (10 mL), saturated aqueous NaHCO3 (10 mL),
brine (10 mL) and dried (Na2SO4). Most of the solvent was
removed under reduced pressure and the precipitated solid was filtered.
The cake was washed with diethyl ether (10 mL) and dried under vacuum to
give 359 (95 mg, 48% yield).

General Procedure for Compounds 297, 298 and 360

[0203] To a solution of intermediate 1-6 (0.84 g, 2.0 mmol) and the
corresponding pyridine carboxylic acid (0.49 g, 4.0 mmol, 2.0 eq) in
anhydrous DMF (25 mL) at room temperature was sequentially added HBTU
(1.52 g, 4.0 mmol, 2.0 eq) and DIEA (3.5 mL, 20 mmol, 10 eq) and the
contents were stirred at room temperature overnight. The reaction mixture
was poured into ice-cold water and the solid obtained was filtered and
dried under vacuum. The free base obtained above was stirred in 2M HCl in
diethyl ether (10 mL), filtered and dried to give the appropriate HCl
salt form of the final compounds.

[0206] To a slurry of 1-8 (100 mg, 0.34 mmol) in anhydrous EtOH (5 mL) was
added a solution of NaOEt in EtOH (2.6 M solution, 0.2 mL, 0.52 mmol) at
room temperature under N2 for 1 h. Then, 1-9 (62 mg, 0.34 mmol) was
added to the mixture and the reaction was heated to reflux for 2 hours.
During that time, the desired product precipitated out. The mixture was
cooled to room temperature and filtered. The solid was washed with EtOH
(10 mL) and diethyl ether (15 mL), and dried under vacuum to give 290a
(53 mg, 39% overall yield).

[0207] To a slurry of 17 (280 mg, 0.704 mmol) in anhydrous EtOH (60 mL)
was added PtO2 (28 mg), and the mixture was hydrogenated at 30 psi
for 3 days. The mixture was filtered through Celite, the filtrate was
concentrated and the resulting residue was recrystallized with
MeOH/diethyl ether (1:4, 5 mL) to give 290 (45 mg, 18% yield).

[0209] To a solution of 1-11 (500 mg, 3.00 mmol) and 2-cyanothioacetamide
(1.0 g, 10.0 mmol) in anhydrous EtOH (36 mL) was added a solution of
NaOEt in EtOH (2.6 M solution, 4.0 mL, 1.04 mmol) at room temperature and
then the mixture was heated to reflux for 1 hour. The mixture was cooled
to room temperature, concentrated and the residue was dissolved in water
(20 mL). Concentrated HCl was added dropwise to adjust the pH to 8-9 when
a solid precipitated out. The precipitate was collected by filtration and
filter cake was washed with water and dried under vacuum to yield 1-12
(212 mg, 34% yield).

[0210] To a slurry of compound 1-12 (150 mg, 0.721 mmol) in anhydrous EtOH
(5 mL) was added 1-5 (216 mg, 0.721 mmol), followed by a solution of
NaOEt in EtOH (2.6 M solution, 0.5 mL, 1.3 mmol) at room temperature
under N2. The reaction was heated to reflux for 2 hours and during
that time, the desired product precipitated out. The mixture was cooled
to room temperature and filtered. The solid was washed with EtOH (2 mL),
diethyl ether (5 mL), and dried under vacuum to give 1-13 (230 mg, 75%
yield).

[0211] To a slurry of compound 1-13 (230 mg, 0.54 mmol) in THF (5 mL) was
added a solution of LiOH in water (1 M solution, 1.35 mL, 1.35 mmol). The
reaction was stirred at room temperature for 2 hours and during that time
the desired product precipitated out. After filtration, the solid was
washed with EtOH (2 mL) and diethyl ether (5 mL), and dried under vacuum
to give 291 (48 mg, 22% yield).

Example 21

Synthesis of Compound 292

##STR00676##

[0213] To a slurry of 1-8 (200 mg, 0.669 mmol) in anhydrous EtOH (10 mL)
was added a solution of NaOEt in EtOH (2.6 M solution, 0.4 mL, 1.04 mmol)
at room temperature under nitrogen for one hour. Then, 1-10 (116 mg,
0.669 mmol) was added to the mixture and the reaction was heated to
reflux for 2 hours. During that time, the desired product precipitated
out. The mixture was cooled to room temperature and filtered. The solid
was washed with EtOH (10 mL), diethyl ether (15 mL), and dried under
vacuum to yield 292 (35 mg, 15% overall yield).

[0216] A solution of 292 (350 mg, 1 eq), TEA (2 ml), and
N-(Boc)-2-bromoethylamine (1 g, 5 eq) in NMP (20 mL) was heated at
100° C. for 16 h. The reaction mixture was cooled to room
temperature, poured into ice water (60 mL), and the solid was filtered
and dried to give the Boc-protected intermediate. This solid dissolved in
10% HCl in MeOH (20 mL) and stirred at room temperature for 3 h. The
volume of the reaction mixture was reduced to 3 mL, the solid was
collected by filtration and washed by diethyl ether (3×3 mL) to
afford product 300 (85 mg, 20% yield) as a light-yellow powder.

[0218] A mixture of 292 (1 g, 1 eq) and TEA (3.33 ml) in anhydrous DCM
(100 mL) was stirred at 0° C., then methyl malonyl chloride (0.833
mL, 3 eq) was added slowly. After stirring at room temperature for 18 h,
DMF (5 mL) was added and the reaction was stirred for an additional 6 h
in attempt to drive to completion. The mixture was concentrated to
dryness, triturated in water (500 mL) for 1 h, filtered, and the solid
was washed by MTBE (3×30 mL). This crude ester intermediate was
purified by silica gel column chromatography using 0-5% MeOH/DCM to give
pure material (385 mg, 31% yield). The hydrolysis reaction was performed
with the purified ester intermediate (386 mg, 1 eq) in 3:1 THF/H2O
(30 mL) and 1M NaOH (3.4 mL, 4.3 eq). The reaction was stirred at room
temperature and then concentrated to dryness. The resulting solid was
collected by filtration, washed by MTBE (3×50 mL), and dried to
give 362 as a light-yellow solid (215 mg, 17% overall yield).

[0233] A solution of 1-25 (500 mg) in 1,4-dioxane was reacted with
bromoacetyl bromide and TEA. After stirring at room temperature for 20
minutes, the reaction mixture was poured into cold diethyl ether, stirred
for 10 min, filtered, washed with diethyl ether and dried in vacuo to
afford 760 mg (quantitative yield) of the bromoacetyl intermediate as the
hydrobromide salt. On 200 mg scale, this bromoacetyl intermediate was
reacted with a methylamine solution (33% wt. solution in EtOH) for 2
hours at room temperature. The reaction mixture was evaporated to dryness
and triturated with DCM to afford pure compound. This material was
treated with 1.25M HCl in MeOH and stirred for 2 hours. Following
evaporation in vacuo and trituration with diethyl ether, 75 mg of
compound 307 was isolated as the HCl salt (44% yield).

[0234] On 200 mg scale, the bromoacetyl intermediate used for the
synthesis of compound 307 was reacted with a 2M dimethylamine solution in
THF for 1 hour at room temperature. The reaction mixture was evaporated
to dryness and treated with 2M HCl in diethyl ether and stirred for 1
hour. The reaction mixture was filtered and triturated with DCM to afford
135 mg of 308 as the HCl salt (79% yield).

[0235] On 150 mg scale, the bromoacetyl intermediate used for the
synthesis of compound 307 was mixed with a 25% trimethylamine in MeOH
solution for 1 hour at room temperature. The reaction mixture was
evaporated to dryness and triturated with DCM to afford 100 mg of 309
(71% yield).

[0245] To a solution of (L)-Cbz-valine (2.5 g, 10 mmol) in anhydrous DMF
(100 mL) at room temperature was added cesium carbonate (3.3 g, 10 mmol)
and the mixture was stirred for 1 h. To the reaction flask was added the
intermediate 1-32 (1 g) and the contents were stirred at room temperature
overnight. The reaction mixture was added to ice-cold water and the
precipitate obtained was filtered, washed with MTBE (2×30 mL) and
dried to afford 316 as a yellow solid (0.5 g).

[0248] To a mixture of 1-34 (3 g, 17.8 mmol, 1 eq) and
2-cyanothioacetamide (2.7 g, 26.8 mmol, 1.5 eq) in ethanol (30 mL) was
added N-methylmorpholine (2.5 mL) and refluxed for 24 h. The reaction
mixture was evaporated in vacuo to afford 7 g of crude 1-35 which was
used in the next step without purification.

[0251] A solution of 1-acetyl-2,4-dimethyl-thiazole (10 g, 64 mmol) in
N,N-dimethylformamide dimethylacetal (100 mL) was refluxed overnight.
GC/MS analysis showed completion. The contents were cooled to room
temperature and poured into ice-cold water. The solid 1-37 obtained (10
g, 80%) was dried and used in the next step as such.

[0252] To a mixture of 1-37 (10 g, 48 mmol) and 2-cyanothioacetamide (10
g, 100 mmol) in EtOH (200 mL) was added NMP (10 mL) and the contents were
heated at 80° C. overnight. The volatiles were removed under
vacuum and the residue was triturated with a 2:1 mixture of hexane/EtOAc
affording the desired intermediate 1-38 (7.2 g, 60% yield) as an orange
solid, which was used in the next step as such.

[0255] Chloroacetonitrile (2.0 g, 26.7 mmol) and
3-(trifluoromethyl)benzenamine (4.20 g, 26.7 mmol) was treated with 4N
HCl in 1,4-dioxane (50 mL). The reaction mixture was stirred at room
temperature overnight. The reaction mixture was concentrated under vacuum
and crude 1-40 was used for next step without further purification.

[0259] To compound 1-41 (5 g, 18 mmol) and 2-chloroacetyl chloride (3.0 g,
27 mmol) in DCM (100 mL) was added a catalytic amount of
tetrabutylammonium hydrosulfate followed by a solution of K2CO3
(5 g, 36 mmol) in water (100 mL). The reaction mixture was stirred at
room temperature for 40 min and the organic portion was isolated and
concentrated which was combined with another reaction product done on the
same scale. The residue was purified via silica gel column chromatography
eluting with 5:1 hexanes/MTBE to give 8 g of 1-42 as a yellowish oil (62%
yield).

[0260] To a solution of compound 1-42 (1.0 g, 2.8 mmol) in DCM was added
10 mL of TFA. The resulting mixture was stirred at room temperature for 2
h and then the solvents were removed. The crude mixture was used for the
next step directly.

[0262] This was a by-product formed resulting from intramolecular
cyclization of the ethyl ester version of compound 326. After performing
base catalyzed hydrolysis of the ester group of this intermediate,
compound 320 was the major product isolated. Note: originally this was an
alternate scheme to synthesize compound 326.

[0264] 3-(trifluoromethyl)-N-methylbenzenamine (3.0 g, 28 mmol) and
2-chloroacetyl chloride (12.6 g, 112 mmol) in 30 mL of DCM was added a
catalytic amount of tetrabutylammonium hydrosulfate, followed by a
solution of K2CO3 (15 g, 112 mmol) in 100 mL of water. The
reaction mixture was stirred at room temperature for 40 min and the DCM
layer was collected and combined with another same scale reaction. The
residue was purified through a silica gel column eluting with 5:1
hexane/MTBE to give 2.7 g of 1-49 as a yellowish oil (38% yield).

[0265] To a mixture of compound 1-49 (2.7 g, 10.7 mmol) and 1-23 (1.5 g,
7.2 mmol) in 20 mL of EtOH was added 15 mL of 21% NaOEt in EtOH. The
reaction mixture was heated for 2 h and then filtered. The solid was
washed 20 mL of EtOH and dried to give 1.8 g of 322 (58% yield).

[0268] To a solution of crude 1-50 (20 g) in anhydrous THF (200 mL) at
0° C. was added dropwise a solution of LiAlH4 (1M solution in
THF, 186 mL, 186 mmol) and the contents were slowly warmed to 70°
C. and refluxed overnight. The contents were cooled to 0° C.,
quenched with the addition of a saturated sodium potassium tartrate
solution and filtered through a pad of Celite. The clear solution was
concentrated and the residue was partitioned between EtOAc (500 mL) and
water (100 mL). The layers were separated and the organic layer was
washed with a saturated NaHCO3 solution, dried (Na2SO4),
filtered and concentrated. The residue obtained was left at high-vacuum
overnight affording the desired intermediate 1-51 (8 g) as a brown oil.

[0270] To a mixture of 1-23 and 1-52 in anhydrous DMF (30 mL) at room
temperature was added K2CO3 (13.8 g, 100 mmol) and the contents
were stirred at 90° C. for 2 days. The contents were cooled to
room temperature and poured into ice-cold water. The solid obtained was
filtered, washed with MTBE (3×50 mL) and dried. The orange solid
obtained (1.5 g) was treated with 4M HCl in dioxane (20 mL) at room
temperature for 5 h and filtered. The orange solid was dried under
high-vacuum affording 323 as the HCl salt (1.2 g).

[0273] To a suspension of compound 1-54 (3.3 g, 13 mmol) in 50 mL of DCM
was added 5 mL of pyridine followed by 3 mL of acetic anhydride. The
reaction mixture was stirred for 2 h and filtered. The solid was
collected and triturated with EtOH (50 mL) at 60° C. for 30
minutes. The solid was collected and dried to give 2.5 g of 1-55 (67%
yield).

[0282] To a solution of 1-59 (2 g, 7.6 mmol), 2-chloroacetyl chloride (3.4
g, 30 mmol), a catalytic amount of tetrabutylammonium hydrosulfate in 40
mL of DCM was added a solution of K2CO3 (4.0 g, 30 mmol) in
water (40 mL). The resulting mixture was stirred at room temperature for
40 min and then the organic layer was collected and concentrated. The
crude mixture was purified through silica gel column chromatography
eluting 4:1 hexanes/MTBE to give 2.8 g of 1-60 as a yellow oil in
quantitative yield.

[0283] To a mixture of 1-60 (2.8 g, 8.3 mmol), 1-23 (1.5 g, 6.9 mmol), and
K2CO3 (11.5 g, 83 mmol) was added 25 mL of DMF. The resulting
mixture was stirred at 50° C. for 2 h and then diluted with water
(1000 mL). Following extraction with EtOAc (1000 mL), the combined
organic layers were dried, filtered, and concentrated. The crude mixture
was triturated with MTBE to give 2 g of 1-61 as a yellow solid (56%
yield).

[0284] To solution of 1-61 (500 mg, 0.96 mmol) in THF was added 40 a 4N
NaOH solution (40 mL). The resulting mixture was stirred at room
temperature overnight. Solvents were removed and the solid was collected,
washed with water (50 mL), THF (5 mL), and dried to give 400 mg of 327 as
yellow solid (85% yield).

Example 36

Synthesis of Compounds 329 and 330

##STR00691##

[0285] Synthesis of 8-oxabicyclo[5.1.0]octan-6-one (1-63)

[0286] To a solution of cyclohept-2-enone (6.0 g, 45.5 mmol) in MeOH (40
mL) was added 13.6 ml of H2O2 at -4° C., followed by 7
mL of 10% NaOH solution. The resulting mixture was stirred at room
temperature for 1 h, diluted with brine (1000 mL), and extracted with
MTBE (2×200 mL). The combined organic layers were dried, filtered,
concentrated and the crude material was purified by silica gel column
chromatography eluting 15:1 hexanes/MTBE to give 5.5 g of 1-63 as a
yellowish oil (96% yield).

Synthesis of Cycloheptane-1,3-dione (1-64)

[0287] To a solution of 1-63 (6.0 g, 47 mmol) in toluene (18 mL) was added
Pd(PPh3)4 (2.7 g, 2.35 mmol) and
1,2-bis(diphenylphosphino)ethane (1.0 g, 2.35 mmol). The reaction was
bubbled with N2 for 10 min, sealed in a 75 mL pressure tube and
heated at 100° C. overnight. The reaction was cooled to room
temperature and the solid was filtered off. The filtrate was collected,
concentrated and purified by silica gel column chromatography eluting
1:10 hexanes/diethyl ether to give 5.0 g of crude product. This material
was distilled to give 3.0 g of 1-64 as a yellowish oil which was used in
the next step directly.

Synthesis of 2-(dimethylaminomethylene)cycloheptane-1,3-dione (1-65)

[0288] A solution of 1-64 (3.0 g, 23.8 mmol) in N,N-dimethylformamide
dimethyl acetal (30 mL) was stirred at room temperature overnight. The
reaction mixture was concentrated in vacuo, the solid was collected and
washed with 1:1 of hexane/diethyl ether (50 mL) to give 3.4 g of 1-65 as
a yellowish solid (79% yield).

[0291] To a solution of 328 (100 mg, 0.23 mmol) in EtOH was added
NaBH4 (100 mg, 2.6 mmol) and the reaction mixture was stirred at
room temperature for 40 min and then quenched with a saturated NH4Cl
solution (20 mL). The solid was collected, washed with water (20 mL), and
dried to give 110 mg of 329 as a yellow solid in quantitative yield.

[0296] For the synthesis of final compounds see the procedure used for
intermediate 1-6. Compound 334 required an additional step involving
hydrolysis of the ester following the cyclization reaction. Note: The
bromoacetamide intermediate used in the final reaction was synthesized
using the same procedure used for the synthesis of 1-24. Please note some
compounds required reduction of the parent nitro moiety to the
corresponding amine and was based upon commercial availability of the
starting materials.

Example 38

Synthesis of Compounds 332, 339 and 345

##STR00693##

[0297] The same experimental procedures used for the compounds above
(i.e., 331, 333, 334, etc.) were used for the synthesis of compounds 332,
339, and 345.

Example 39

Synthesis of Compound 346

##STR00694##

[0298] Synthesis of p-tolyl 4-nitrobenzenesulfonate (1-74)

[0299] To a solution of compound 1-73 (4 g, 37 mmol), pyridine (4.5 mL)
and THF (50 mL) was added a solution of p-cresol (9.8 g) in THF (25 mL)
slowly over 10 min at 0° C. The reaction mixture was allowed to
reach ambient temp and then heated to 65° C. for 48 h. The
reaction was stopped by adding a saturated aqueous NH4Cl solution
and extracted with EtOAc. The organic layer was washed with brine, dried
over Na2SO4, filtered and concentrated under vacuum to give a
residue. The residue was purified by silica gel column chromatography
eluting with 0-50% EtOAc/Hexanes to give 7.7 g of compound 1-74.

Synthesis of p-tolyl 4-aminobenzenesulfonate (1-75)

[0300] To a mixture of compound 1-74 (2 g, 6.8 mmol, 1.0 equiv.) in EtOH
(40 mL) was added a solution of NH4Cl (1.5 g, 27 mmol, 4.0 equiv.)
in 10 mL of water followed by iron (1.5 g, 27 mmol, 4.0 equiv.). The
reaction mixture was heated to 80° C. for 20 min, cooled to
ambient temp, filtered through a pad of Celite, and then washed with MeOH
and DCM. The combined filtrates were concentrated under vacuum and
extracted with DCM. The organic portion was washed with water, dried
(Na2SO4), filtered and concentrated under vacuum to give crude
material. The crude product was purified by silica gel column
chromatography to give 1.1 g of compound 1-75 (61% yield).

Synthesis of p-tolyl 4-[(2-bromoacetyl)amino]benzenesulfonate (1-76)

[0301] To a solution of compound 1-75 (1.1 g, 4.2 mmol, 1.0 equiv.) in THF
(100 mL) was added NaHCO3 (5.3 g, 6.3 mmol, 1.5 equiv.) and
bromoacetyl bromide (0.44 mL, 5.02 mmol, 1.2 equiv.) at 0° C. The
reaction mixture was warmed to ambient temp and stirred for 16 h. The
reaction mixture was filtered through a pad of Celite, washed with DCM,
and the combined filtrates were concentrated under vacuum to give crude
compound 1-76. This material was carried to next step without further
purification.

[0303] A mixture of compound 1-77 (425 mg), 10 mL of 20% NaOH in water and
MeOH (10 mL) was heated to 80° C. for 14 h. The mixture was cooled
to ambient temperature and the solids were removed by filtration, washed
with water, DCM, hexanes and dried under vacuum. The solids were
suspended in water (5 mL) and acidified with 3N HCl to adjust the pH to
2-3 and stirred for 30 min. The solids were filtered, washed with water,
DCM and hexanes. The solids were dried under vacuum at 35° C. for
14 h to give 210 mg of 346 (59% overall yield).

Example 40

Synthesis of Compounds 350, 354 and 355

##STR00695##

[0305] The same experimental procedures used for the compound 327 were
used for the synthesis of compounds 350, 354, and 355.

[0339] All references cited herein are herein incorporated by reference in
their entirety for all purposes.

[0340] The invention has been described in terms of preferred embodiments
thereof, but is more broadly applicable as will be understood by those
skilled in the art. The scope of the invention is only limited by the
following claims.

Patent applications by Chelsea M. Byrd, Corvallis, OR US

Patent applications by Dennis E. Hruby, Albany, OR US

Patent applications by Dongcheng Dai, Corvallis, OR US

Patent applications by Shanthakumar R. Tyavanagimatt, Corvallis, OR US